Intermittent fasting can benefit breast cancer patients undergoing chemotherapy or immunotherapy.However,it is still uncertain how to select immunotherapy drugs to combine with intermittent fasting.Herein we observed ...Intermittent fasting can benefit breast cancer patients undergoing chemotherapy or immunotherapy.However,it is still uncertain how to select immunotherapy drugs to combine with intermittent fasting.Herein we observed that two cycles of fasting treatment significantly inhibited breast tumor growth and lung tissue metastasis,as well as prolonged overall survival in mice bearing 4T1 and 4T07 breast cancer.During this process,both the immunosuppressive monocytic-(M-)and granulocytic-(G-)myeloid-derived suppressor cell(MDSC)decreased,accompanied by an increase in interleukin(IL)7R^(+)and granzyme B^(+)T cells in the tumor microenvironment.Interestingly,we observed that Ly6G^(low)G-MDSC sharply decreased after fasting treatment,and the cell surface markers and protein mass spectrometry data showed potential therapeutic targets.Mechanistic investigation revealed that glucose metabolism restriction suppressed the splenic granulocytemonocyte progenitor and the generation of colony-stimulating factors and IL-6,which both contributed to the accumulation of G-MDSC.On the other hand,glucose metabolism restriction can directly induce the apoptosis of Ly6G^(low)G-MDSC,but not Ly6G^(high)subsets.In summary,these results suggest that glucose metabolism restriction induced by fasting treatment attenuates the immune-suppressive milieu and enhances the activation of CD3^(+)T cells,providing potential solutions for enhancing immune-based cancer interventions.展开更多
Ionizing radiations are tools in diagnosis and treatment of diseases. Leukopenia from exposure to ionizing radiation has been reported. Due to their radiosensitivity, leukocytes are a biological model to analyze cell ...Ionizing radiations are tools in diagnosis and treatment of diseases. Leukopenia from exposure to ionizing radiation has been reported. Due to their radiosensitivity, leukocytes are a biological model to analyze cell damage. Therefore, cell viability, DNA damage, and Hsp70 and p53 expression in human leukocytes exposed to low-dose gamma radiation fields from a <sup>137</sup>Cs source were evaluated. A decrease in cell viability, DNA damage and an increase in the expression of Hsp70 and p53 proportional to the radiation dose received was found, which was 0.2, 0.4, 0.6, 0.8 and 1.0 mGy.展开更多
Salinity is one of the most severe abiotic stresses for crop production.The present study investigates the salinityinduced modulation in growth indicators,morphology and movement of stomata,photosynthetic pigments,act...Salinity is one of the most severe abiotic stresses for crop production.The present study investigates the salinityinduced modulation in growth indicators,morphology and movement of stomata,photosynthetic pigments,activity of carbonic anhydrase as well as nitrate reductase,and antioxidant systems in two varieties of chickpea(Pusa-BG5023,and Pusa-BGD72).On 20^(th) day of sowing,plants were treated with varying levels of NaCl(0,50,100,150 and 200 mM)followed by sampling on 45 days of sowing.Recorded observations on both the varieties reveal that salt stress leads to a significant decline in growth,dry biomass,leaf area,photosynthetic pigments,protein content,stomatal behavior,cell viability,activity of nitrate reductase and carbonic anhydrase with the rise in the concentration of salt.However,quantitatively these changes were less in Pusa-BG5023 as compared to Pusa-BGD72.Furthermore,salinity-induced oxidative stress enhanced malondialdehyde content,superoxide radicals,foliar proline content,and the enzymatic activities of superoxide dismutase,catalase,and peroxidase.The variety Pusa-BGD72 was found more sensitive than Pusa-BG5023 to salt stress.Out of different graded concentrations(50,100,150 and 200 mM)of sodium chloride,50 mM was least toxic,and 200 mM was most damaging.The differential behavior of these two varieties measured in terms of stomatal behavior,cell viability,photosynthetic pigments,and antioxidant defense system can be used as prospective indicators for selection of chickpea plants for salt tolerance and sensitivity.展开更多
The hardness,wettability,and electrochemical properties of Ti6Al4V alloy surfaces treated with anodic oxidation and plasma oxidation as well as the viabilities of the different cell lines on the obtained surfaces were...The hardness,wettability,and electrochemical properties of Ti6Al4V alloy surfaces treated with anodic oxidation and plasma oxidation as well as the viabilities of the different cell lines on the obtained surfaces were investigated.The anodic oxidation was performed for 10 min under 100 V potential,and it resulted in a 0.95μm thick nanoporous anatase-TiO2 structure.On the other hand,plasma oxidation was carried out at 650℃ for 1 h and resulted in a dense rutile-TiO2 structure with a thickness of 1.2μm.While a hardness of HV0.025823 and roughness of^220 nm were obtained by plasma oxidation,those obtained by anodic oxidation were HV0.025512 and^130 nm,respectively.The anodic oxidation process created a more hydrophilic surface with a contact angle of 87.2°.Both oxidation processes produced similar properties in terms of corrosion behavior and showed better resistance than the as-received state in a certain range of potential.Moreover,the surface treatments led to no significant change in the protein adsorption levels,which indicates that the difference in viability between the osteoblast and fibroblast cells was not due to the difference in surface protein adsorption.Given all the factors,the surfaces obtained by anodic oxidation treatment revealed higher cell viability than those obtained by plasma oxidation(p=0.05).展开更多
Although nanocomposites have recently attracted special interest in the tissue engineering area,due to their potential to reinforce scaffolds for hard tissues applications,a number of variables must be set prior to an...Although nanocomposites have recently attracted special interest in the tissue engineering area,due to their potential to reinforce scaffolds for hard tissues applications,a number of variables must be set prior to any clinical application.This manuscript addresses the evaluation of thermo-mechanical properties and of cell proliferation of cellulose nanocrystals(CNC),poly(butylene adipate-co-terephthalate)(PBAT),poly(ε-caprolactone)(PCL)films and their bionanocomposites with 2 wt% of CNC obtained by casting technique.Cellulose nanocrystals extracted from Balsa wood by acid hydrolysis were used as a reinforcing phase in PBAT and PCL matrix films.The films and pure CNC at different concentrations were cultured with osteoblasts MG-63 and the cell proliferation was assessed by AlamarBlue?assay.The thermal-mechanical properties of the films were evaluated by dynamic-mechanical thermal analysis(DMTA).It was found by DMTA that the CNC acted as reinforcing agent.The addition of CNCs in the PBAT and PCL matrices induced higher storage moduli due to the reinforcement effects of CNCs.The cell viability results showed that neat CNC favored osteoblast proliferation and both PBAT and PCL films incorporated with CNC were biocompatible and supported cell proliferation along time.The nature of the polymeric matrix or the presence of CNC practically did not affect the cell proliferation,confirming they have no in vitro toxicity.Such features make cellulose nanocrystals a suitable candidate for the reinforcement of biodegradable scaffolds for tissue engineering and biomedical applications.展开更多
Cadmium(Cd)is one of the most widespread and toxic heavy metals to plants.Extracellular ATP(exATP)is thought to be an extracellular effector in regulating the physiological responses of plant cells to environmental st...Cadmium(Cd)is one of the most widespread and toxic heavy metals to plants.Extracellular ATP(exATP)is thought to be an extracellular effector in regulating the physiological responses of plant cells to environmental stresses.However,the function of exATP in Cd-stressed plant cells is much unknown.The present work showed that treating tobacco(Nicotiana tabacum L.cv.Bright Yellow-2)cell-suspension cultures with exogenous CdCl2 reduced the cell viability,exATP level,and Mg content.However,the production of reactive oxygen species(ROS),Cd content,and electrolyte leakage of the cells were enhanced by exogenous CdCl2.When the Cd-induced accumulation of ROS was decreased by the supplement with DMTU(dimethylthiourea,a scavenger of ROS),the Cd-induced increases of the electrolyte leakage and Cd content were alleviated,and the Cd-induced reductions of cell viability were partly rescued,suggesting that Cd-induced reduction of cell viability could be related to the ROS accumulation.Under the condition of Cd stress,when the reduction of exATP level was partly rescued by exogenous ATP(20μM),the increases of ROS production,electrolyte leakage,and Cd content were attenuated,and the reduction of cell viability was also alleviated.These observations indicate that exATP can regulate the cell viability in the Cd–stressed plant cells possibly by an ROS-associated mechanism.展开更多
Cellular radiosensitivity is directly correlated with the mechanism of DNA repair, in which p53 protein plays a major role. In this context, this study correlated cell death with p53 expression in lymphocytes irradiat...Cellular radiosensitivity is directly correlated with the mechanism of DNA repair, in which p53 protein plays a major role. In this context, this study correlated cell death with p53 expression in lymphocytes irradiated in vitro with different doses of gammaradiation. For this, peripheral blood samples were collected from 10 healthy subjects. Each sample was divided in aliquots and, separately, irradiated with doses of 0,5;2 and 4 Gy. After this, peripheral blood mononuclear cells (PBMCs) were isolated and cultivated during 72 hours in 5% CO2 at 37oC without mitogen stimulation. The expression of p53 protein was evaluated by flow cytometry. In parallel, cell viability was determined by trypan blue staining. Statistical analysis was performed us-ing analysis of variance (ANOVA), differences were considered as statistically significant when p < 0.05. The results showed an increase of p53 expression with the absorbed dose, which was proportional to cell death, suggesting that p53 can be used as bioindicator of individual radiosensitivity.展开更多
·AIM:To transduce recombinant human platelet-derived growth factor B(PDGF-B) gene adeno-associated virus(AAV) to in vitro cultured cat corneal endothelial cell(CEC) and observe the effect of the expressed PDGF-BB...·AIM:To transduce recombinant human platelet-derived growth factor B(PDGF-B) gene adeno-associated virus(AAV) to in vitro cultured cat corneal endothelial cell(CEC) and observe the effect of the expressed PDGF-BB protein on the viability of cat CEC.·METHODS:Cat cornea endothelium was torn under microscope and rapidly cultivated in DMEM to form single layer CEC and the passage 2 endothelial cells were used in this study. The recombinant human PDGF-B gene AAV was constructed and transduced into cat CEC directly. Three groups were as following:blank control group, AAV control group and recombinant AAV group. At 24 hours, 48 hours, and 5 days after transduction, total RNA was extracted from the CEC by Trizol and the expression of PDGF-B gene was detected by fluorescence quantitative polymerase chain reaction. Viability of the transduced CEC was detected at 48 hours after transduction by MTT assay. Cell morphology was observed under inverted phase contrast microscope.·RESULTS:With the torn endothelium culture technique, we rapidly got single layer cat CEC. At 24 hours, 48 hours and 5 days after transduction, fluorescence quantitative polymerase chain reaction showed there was no significant difference of the expressed PDGF-B gene mRNA between blank control group and AAV control group (P >0.05). In contrast, there were significant differences between two control groups and recombinant AAV group (P <0.05). MTT assay showed that in recombinant AAV group, the expressed PDGF-BB protein could promote the viability of cat CEC. Morphology observation showed at 48 hours after transduction, cells in CEC-AAV-PDGF-B group proliferated into bigger scales in regular triangle to hexagon shape with distinct boundary, while the number of cells was significantly less in the two control groups.·CONCLUSION:The recombinant AAV-PDGF-B expresses biological active PDGF-BB protein in cat CEC, which promotes the viability and proliferation of cells.展开更多
AIM:To investigate the effect of aspirin on neuroendocrine tumor(NET)cell growth and signaling in vitro.METHODS:Human pancreatic BON1,bronchopulmonary NCI-H727 and midgut GOT1 neuroendocrine tumor cells were treated w...AIM:To investigate the effect of aspirin on neuroendocrine tumor(NET)cell growth and signaling in vitro.METHODS:Human pancreatic BON1,bronchopulmonary NCI-H727 and midgut GOT1 neuroendocrine tumor cells were treated with different concentrations of aspirin(from 0.001 to 5 mmol/L),and the resulting effects on metabolic activity/cell proliferation were measured using cell proliferation assays and SYBR-DNAlabeling after 72,144 and 216 h of incubation.The effects of aspirin on the expression and phosphorylation of several critical proteins that are involved in the most common intracellular growth factor signaling pathways(especially Akt protein kinase B)and mammalian target of rapamycin(mTOR)were determined by Western blot analyses.Propidium iodide staining and flow cytometry were used to evaluate changes in cell cycle distribution and apoptosis.Statistical analysis was performed using a 2-tailed Student’s t-test to evaluate the proliferation assays and cell cycle analyses.The results are expressed as the mean±SD of 3 or 4 independently performed experiments.Statistical significance was set at P<0.05.RESULTS:Treatment with aspirin suppressed the viability/proliferation of BON1,NCI-H727 and GOT1 cells in a time-and dose-dependent manner.Significant effects were observed at starting doses of 0.5-1 mmol/L and peaked at 5 mmol/L.For instance,after treatment with 1 mmol/L aspirin for 144 h,the viability of pancreatic BON1 cells decreased to 66%±13%(P<0.05),the viability of bronchopulmonary NCI-H727 cells decreased to 53%±8%(P<0.01)and the viability of midgut GOT1 cells decreased to 89%±6%(P<0.01).These effects were associated with a decreased entry into the S phase,the induction of the cyclin-dependent kinase inhibitor p21 and reduced expression of cyclindependent kinase 4 and cyclin D3.Aspirin suppressed mTOR downstream signaling,evidenced by the reduced phosphorylation of the mTOR substrates 4E binding protein 1,serine/threonine kinase P70S6K and S6 ribosomal protein and inhibited glycogen synthase kinase3 activity.We observed the(compensatory)activation of tuberous sclerosis 2,the serine/threonine specific protein kinase AKT and extracellular signal-regulated kinases.CONCLUSION:Aspirin demonstrates promising anticancer properties for NETs in vitro.Further preclinical and clinical studies are needed.展开更多
An argon atmospheric pressure plasma jet was employed to treat L929 murine fibroblasts cultured in vitro.Experimental results showed that,compared with the control cells,the treatment of fibroblasts with 15 s of plasm...An argon atmospheric pressure plasma jet was employed to treat L929 murine fibroblasts cultured in vitro.Experimental results showed that,compared with the control cells,the treatment of fibroblasts with 15 s of plasma led to a significant increase of cell viability and collagen synthesis,while the treatment of 25 s plasma resulted in a remarkable decrease.Exploration of related mechanisms suggested that cold plasma could up-regulate Cyclin D1 gene expression and down-regulate p27 gene expression at a low dose,while it could down-regulate Cyclin D1 expression and up-regulate p27 expression at a higher dose,thus altering the cell cycle progression,and then affecting cell viability and collagen synthesis of fibroblasts.展开更多
AIM: To analyze the concentration-dependent effects of autologous serum(AS) and fetal bovine serum(FBS) on human corneal epithelial cell(HCEC) viability, migration and proliferation.METHODS: AS was prepared from 13 pa...AIM: To analyze the concentration-dependent effects of autologous serum(AS) and fetal bovine serum(FBS) on human corneal epithelial cell(HCEC) viability, migration and proliferation.METHODS: AS was prepared from 13 patients with nonhealing epithelial defects Dulbecco's modified eagle medium/Ham's F12(DMEM/F12) with 5% FBS, 0.5% dimethyl sulphoxide(DMSO), 10 ng/m L human epidermal growth factor, 1% insulin-transferrin-selenium, then were incubated in serum media: DMEM/F12 supplemented by 5%, 10%, 15% or 30% AS or FBS. HCEC viability was analyzed using cell proliferation kit XTT, migration using a wound healing assay, proliferation by the cell proliferation enzyme-linked immunosorbent assay(ELISA) Brd U kit. Statistical analysis was performed using the generalized linear model, the values at 30% AS or 30% FBS were used as the baselines.RESULTS: HCEC viability was the highest at 30% AS or 15% FBS and the lowest at 10% AS or 30% FBS application. HCEC migration was the quickest through 30% AS or 30% FBS and the slowest through 5% AS or 5% FBS concentrations. Proliferation was the most increased through 15% AS or 5% FBS and the least increased through 30% AS or 30% FBS concentrations. HCEC viability at 10% and 15% AS was significantly worse(P=0.001, P=0.023) compared to baseline and significantly better at 15% FBS(P=0.003) concentrations. HCEC migration was significantly worse(P≤0.007) and HCEC proliferation significantly better(P<0.001) in all concentration groups compared to baseline.CONCLUSION: For the best viability of HCEC 30% AS or 15% FBS, for HCEC migration 30% AS or 30% FBS, for proliferation 15% AS or 5% FBS should be used. Therefore, we suggest the use of 30% AS in clinical practice.展开更多
Background: Mammalian ovaries contain follicles containing an oocyte enclosed by layers of granulosa cells (GC). Follicle growth and oocyte maturation are largely dependent on GC numbers and viability, but there is no...Background: Mammalian ovaries contain follicles containing an oocyte enclosed by layers of granulosa cells (GC). Follicle growth and oocyte maturation are largely dependent on GC numbers and viability, but there is no established, reliable method for assessing the number of viable GC within an isolated follicle. Methods: Centrifugation conditions and the Trypan Blue (TB) Exclusion assay were optimised for low cell densities compatible with the numbers of GC in follicles. Mouse ovarian follicles were disaggregated to produce a single cell suspension of GC which were examined by TB (n = 4), but also by crystal violet assay in a 96-well plate format after 24 h in vitro (n = 3). GC viability in vitro was characterised further by using enzyme-linked immunoassays to quantify GC production of anti-Mullerian hormone (AMH) and estrogen. Results: The centrifugation and low cell density TB protocol could accurately measure the viability of 78 GC in 10 μL, with an intra-assay coefficient of variation (CoV) 22%, and inter-assay CoV 7%. The best follicle disaggregation method (30 min 37°C exposure to 2 mg/mL collagenase prior to 30 min exposure to 0.025% hyaluronidase) yielded (656 ±87) GC per antral follicle of which 82% ±5% were viable. Culturing 312 - 20,000 GC per well for 24 hours and assessing viability by crystal violet assay generated a linear correlation between OD value and viable GC number (R2 = 0.98) and estrogen concentration per well (R2 = 0.92). 20,000 GC per well produced 143 ±16 pg/mL estrogen during 24 hours in vitro, but no detectable AMH. Conclusion: This is the first report describing the isolation of viable, estrogen-producing GC from murine follicles, and their subsequent culture. These procedures are transferrable to other species including humans and can be applied to screening the reproductive toxicity of pharmaceutical agents.展开更多
Background: Despite the popularity of autologous fat transfer applications, high resorption rates, and consequential volume loss, have been reported. Viable adipocyte content has been defined as a key determinant of f...Background: Despite the popularity of autologous fat transfer applications, high resorption rates, and consequential volume loss, have been reported. Viable adipocyte content has been defined as a key determinant of fat transfer longevity. Moreover, traces of blood, free oil fat and fibrotic tissue accelerate adipocyte degradation. Objective: To compare the effectiveness of a 1470 nm, radial emitting laser-assisted liposection device to a mechanical liposection device in maintaining adipocyte viability in fat tissue harvests. Methods: Bilateral subcutaneous adipose tissue samples were harvested from ten female patients. Fat was harvested from one side using the LipoLife laser-assisted liposuction device and from the other side with a Byron mechanical aspirator. Samples were visually analyzed and blood:fat ratios and cell viability were determined. Results: Laser-harvested samples separated into two distinct phases, with a negligible blood phase at the bottom (1.1%) and a significant adipose phase at the top (98.9%), containing small, uniform-sized cells, of which 95.7% ± 2.7% proved viable. Mechanically harvested samples separated into blood (18%), adipose (60%) and lipid (22%) phases. The adipose phase contained significant amounts of connective tissue, large adipose tissue fragments, large oil droplets and a mean 79.7% ± 18.3% viable adipocytes. Conclusions: Laser liposuctioning was superior to mechanical liposuctioning, providing both higher cell viability and enhanced sample quality. The 1470 nm diode laser bears the potential of improving long-term clinical outcomes of fat transfer procedures. Improved purity of the harvested sample and heightened preadipocyte content are projected to provide for extended graft longevity.展开更多
Vaccination with tumor-antigen pulsed, monocyte-derived dendritic cells (DCs) has emerged as a promising strategy in cancer immunotherapy. The standard DC maturation cocktail consists of a combination of tumor necrosi...Vaccination with tumor-antigen pulsed, monocyte-derived dendritic cells (DCs) has emerged as a promising strategy in cancer immunotherapy. The standard DC maturation cocktail consists of a combination of tumor necrosis factor-α (TNF-α)/interleukin (IL)-1β/IL-6 and prostaglandin E2 (PGE2) for generation of standard DCs (sDCs). In order to improve IL-12p70 production and cytotoxic T-lymphocyte (CTL) induction, a novel cocktail composed of TNF-α/IL-1β/ interferon (IFN)-α/IFN-γ and polyinosinic:polycytidylic acid (Poly-I:C) has been introduced to generate so-called α-Type-1 polarized DCs (αDC1s). We and others have previously performed a comprehensive comparison of sDCs and αDC1s. Here we demonstrate that the viability of αDC1s is lowered compared to sDCs and that DC apoptosis is mediated by Poly-I:C. We speculated that activation of protein kinase R (PKR) could mediate the observed apoptosis, but despite significantly higher PKR expression in αDC1s compared to sDCs and induction of active threonine (Thr)446 autophosphorylation of PKR in αDC1s, Poly-I:C did not influence total PKR expression or autophosporylation, indicating PKR-independent Poly-I:C-induced DC apoptosis.展开更多
In the present study, PC12 cells induced by 6-hydroxydopamine as a model of Parkinson's Disease, were used to investigate the protective effects of bone marrow-derived mesenchymal stem cells bone marrow-derived me...In the present study, PC12 cells induced by 6-hydroxydopamine as a model of Parkinson's Disease, were used to investigate the protective effects of bone marrow-derived mesenchymal stem cells bone marrow-derived mesenchymal stem cells against 6-hydroxydopamine-induced neurotoxicity and to verify whether the mechanism of action relates to abnormal α-synuclein accumulation in cells. Results showed that co-culture with bone marrow-derived mesenchymal stem cells enhanced PC12 cell viability and dopamine secretion in a cell dose-dependent manner. MitoLight staining was used to confirm that PC12 cells co-cultured with bone marrow-derived mesenchymal stem cells demonstrate reduced levels of cell apoptosis. Immunocytochemistry and western blot analysis found the quantity of α-synuclein accumulation was significantly reduced in PC12 cell and bone marrow-derived mesenchymal stem cell co-cultures. These results indicate that bone marrow-derived mesenchymal stem cells can attenuate 6-hydroxydopamine-induced cytotoxicity by reducing abnormal α-synuclein accumulation in PC12 cells.展开更多
It is well known that α-synuclein (αS) plays an important role in the pathogenesis of Parkinson’s disease (PD). Moreover, oxidative stress is also thought to be an important factor in PD due to induction of dopamin...It is well known that α-synuclein (αS) plays an important role in the pathogenesis of Parkinson’s disease (PD). Moreover, oxidative stress is also thought to be an important factor in PD due to induction of dopaminergic neuronal cell death by free radicals and enhancement of αS fibrillation by oxidized stress. In the present study, to clarify the role of glutathione (GSH), an intracellular antioxidant, on the molecular mechanism of αS-induced cell injury, we examined the effects of L-buthionine-SR-sulfoximine (BSO), a GSH synthase inhibitor, with or without N-acetyl-L-cysteine (NAC), a source of GSH, on αS-induced cell injury in human neuroblastoma SH-SY5Y cells. Treatment with BSO significantly reduced the cell viability of both empty-vector- and αS-transfected SH-SY5Y cells in a dose-dependent manner (p < 0.01), although the ratio of αS-induced reduction of cell viability in α-syn-transfected cells was much greater than that in empty-vector-transfected cells. Moreover, BSO significantly reduced the intracellular total GSH level in both types of transformant cells. However, NAC significantly prevented BSO-induced reduction of both cell viability and GSH level in the αS-transfected cells. These findings suggest that GSH plays an important role in αS-induced cell injury by reducing cell viability.展开更多
Magnetic resonance image (MRI) systems with a much higher magnetic flux densitywere developed and applied for potential use in medical diagnostic. Recently, much attention hasbeen paid to the biological effects of sta...Magnetic resonance image (MRI) systems with a much higher magnetic flux densitywere developed and applied for potential use in medical diagnostic. Recently, much attention hasbeen paid to the biological effects of static, strong magnetic fields (SMF). With the 13 T SMFfacility in the Institute of Plasma Physics, Chinese Academy of Sciences, the present study focusedon the cellular effects of the SMF with 13 T on the cell viability and the cell cycle distributionin immortalized hamster cells, such as human-hamster hybrid (A_L) cells, Chinese hamster ovary(CHO) cells, DNA double-strand break repair deficient mutant (XRS-5) cells, and human primaryskin fibroblasts (AG1522) cells. It was found that the exposure of 13 T SMF had less effect onthe colony formation in either nonsynchronized or synchronized A_L cells. Moreover, as comparedto non-exposed groups, there were slight differences in the cell cycle distribution no matter ineither synchronized or nonsynchronized immortalized hamster cells after exposure to 13 T SMF.However, it should be noted that the percentage of exposed AG1522 cells at G0/G1 phase wasdecreased by 10% as compared to the controls. Our data indicated that although 13 T SMF hadminimal effects in immortalized hamster cells, the cell cycle distribution was slightly modified bySMF in human primary fibroblasts.展开更多
The traditional MTT assay requires destructive analyses and is not convenient in continuous monitory of cell viability. However, a new cell model was developed in this research, by using the oxidation-reduction (redox...The traditional MTT assay requires destructive analyses and is not convenient in continuous monitory of cell viability. However, a new cell model was developed in this research, by using the oxidation-reduction (redox) indicator alamarblue? instead of the MTT assay. The alamarblue?does no harm to cells and provide a much more safe and convenient methods of measurement. Firstly, cell apoptosis was induced by different concentration of 6-OHDA, then the cell viability was tested by the alamarblue?at a serial of time points. Finally, the optimism cells density, 6-OHDA concentration and testing time point were gained to set a wonderful SH-SY5Y cell model in our research. And when it is applied in the study of neuroprotection effects of NAC, GSH and Catalase, the new model reveals undeniable advantages.展开更多
AIM:To investigate the impact of adipose-derived mesenchymal stem cells(ADSCs) on cell viability and extracellular matrix(ECM) synthesis of corneal stromal cells(CSCs). METHODS:ADSCs and CSCs were obtained from the co...AIM:To investigate the impact of adipose-derived mesenchymal stem cells(ADSCs) on cell viability and extracellular matrix(ECM) synthesis of corneal stromal cells(CSCs). METHODS:ADSCs and CSCs were obtained from the corneas of New Zealand white rabbits and indirectly cocultured in vitro. The proliferative capacity of CSCs in the different groups was assessed by CCK-8 assays. Annexin V-fluorescein isothiocyanate(FITC)/proliferation indices(PI) assays were used to detect the apoptosis of CSCs. The expression levels of matrix metalloproteinase(MMP), such as MMP1, MMP2, MMP9, and collagens were also evaluated by Western blot. RESULTS:ADSCs significantly promoted proliferation and invasion of CSCs in the indirect co-culture assays. The co-cultural group displayed much higher ability of proliferation, especially under the co-culture conditions of ADSCs for 3d, compared with that CSCs cultured alone. The PI of CSCs in the co-culture system were increased approximately 3-8-fold compared with the control group. A significant change was observed in the proportions of cells at apoptosis(early and late) between the negative control group(6.34% and 2.06%) and the ADCSs-treated group(4.69% and 1.59%). The expression levels of MMPs were down regulated in the co-culture models. Compared with the control group, the decrease intensities of MMP-1, MMP-2 and MMP-9 in CSCs/ADSCs group were observed, 3.90-fold, 1.09-fold and 3.03-fold, respectively. However, the increase intensities of collagen type(I, II, III, IV, and V) in CSCs were observed in CSCs/ADSCs group, 3.47-fold,4.30-fold, 2.35-fold, 2.55-fold and 2.43-fold, respectively, compared to that in the control group. The expressions of aldehyde dehydrogenase and fibronectin in CSCs were upregulated in the co-culture models.CONCLUSION:ADSCs play a promotive role in CSCs' growth and invasion, which may be partially associated with MMPs decrease and collagens increase, resulting in a positive participation in the plasticity and ECM synthesis of CSCs. This provided a new insight into the extensive role of ADSCs in CSCs and a potential molecular target for corneal therapy.展开更多
基金supported by the Postdoctoral Research Funds of Hebei Medical University(30705010016-3759)Natural Science Foundation of China(32272328)+4 种基金Natural Science Foundation of Hebei Province(B2022321001)National Key Research Project of Hebei Province(20375502D)Postdoctoral Research Project of Hebei Province(B2022003031)Science and Technology Research Program of Hebei Provincial Colleges(QN2023229)Hebei Provincial Key Laboratory of Nutrition and Health(2023YDYY-KF05)。
文摘Intermittent fasting can benefit breast cancer patients undergoing chemotherapy or immunotherapy.However,it is still uncertain how to select immunotherapy drugs to combine with intermittent fasting.Herein we observed that two cycles of fasting treatment significantly inhibited breast tumor growth and lung tissue metastasis,as well as prolonged overall survival in mice bearing 4T1 and 4T07 breast cancer.During this process,both the immunosuppressive monocytic-(M-)and granulocytic-(G-)myeloid-derived suppressor cell(MDSC)decreased,accompanied by an increase in interleukin(IL)7R^(+)and granzyme B^(+)T cells in the tumor microenvironment.Interestingly,we observed that Ly6G^(low)G-MDSC sharply decreased after fasting treatment,and the cell surface markers and protein mass spectrometry data showed potential therapeutic targets.Mechanistic investigation revealed that glucose metabolism restriction suppressed the splenic granulocytemonocyte progenitor and the generation of colony-stimulating factors and IL-6,which both contributed to the accumulation of G-MDSC.On the other hand,glucose metabolism restriction can directly induce the apoptosis of Ly6G^(low)G-MDSC,but not Ly6G^(high)subsets.In summary,these results suggest that glucose metabolism restriction induced by fasting treatment attenuates the immune-suppressive milieu and enhances the activation of CD3^(+)T cells,providing potential solutions for enhancing immune-based cancer interventions.
文摘Ionizing radiations are tools in diagnosis and treatment of diseases. Leukopenia from exposure to ionizing radiation has been reported. Due to their radiosensitivity, leukocytes are a biological model to analyze cell damage. Therefore, cell viability, DNA damage, and Hsp70 and p53 expression in human leukocytes exposed to low-dose gamma radiation fields from a <sup>137</sup>Cs source were evaluated. A decrease in cell viability, DNA damage and an increase in the expression of Hsp70 and p53 proportional to the radiation dose received was found, which was 0.2, 0.4, 0.6, 0.8 and 1.0 mGy.
文摘Salinity is one of the most severe abiotic stresses for crop production.The present study investigates the salinityinduced modulation in growth indicators,morphology and movement of stomata,photosynthetic pigments,activity of carbonic anhydrase as well as nitrate reductase,and antioxidant systems in two varieties of chickpea(Pusa-BG5023,and Pusa-BGD72).On 20^(th) day of sowing,plants were treated with varying levels of NaCl(0,50,100,150 and 200 mM)followed by sampling on 45 days of sowing.Recorded observations on both the varieties reveal that salt stress leads to a significant decline in growth,dry biomass,leaf area,photosynthetic pigments,protein content,stomatal behavior,cell viability,activity of nitrate reductase and carbonic anhydrase with the rise in the concentration of salt.However,quantitatively these changes were less in Pusa-BG5023 as compared to Pusa-BGD72.Furthermore,salinity-induced oxidative stress enhanced malondialdehyde content,superoxide radicals,foliar proline content,and the enzymatic activities of superoxide dismutase,catalase,and peroxidase.The variety Pusa-BGD72 was found more sensitive than Pusa-BG5023 to salt stress.Out of different graded concentrations(50,100,150 and 200 mM)of sodium chloride,50 mM was least toxic,and 200 mM was most damaging.The differential behavior of these two varieties measured in terms of stomatal behavior,cell viability,photosynthetic pigments,and antioxidant defense system can be used as prospective indicators for selection of chickpea plants for salt tolerance and sensitivity.
基金This work was financially supported by Erzincan Binali Yıldırım University Research Fund(No.FBA-2018-547).
文摘The hardness,wettability,and electrochemical properties of Ti6Al4V alloy surfaces treated with anodic oxidation and plasma oxidation as well as the viabilities of the different cell lines on the obtained surfaces were investigated.The anodic oxidation was performed for 10 min under 100 V potential,and it resulted in a 0.95μm thick nanoporous anatase-TiO2 structure.On the other hand,plasma oxidation was carried out at 650℃ for 1 h and resulted in a dense rutile-TiO2 structure with a thickness of 1.2μm.While a hardness of HV0.025823 and roughness of^220 nm were obtained by plasma oxidation,those obtained by anodic oxidation were HV0.025512 and^130 nm,respectively.The anodic oxidation process created a more hydrophilic surface with a contact angle of 87.2°.Both oxidation processes produced similar properties in terms of corrosion behavior and showed better resistance than the as-received state in a certain range of potential.Moreover,the surface treatments led to no significant change in the protein adsorption levels,which indicates that the difference in viability between the osteoblast and fibroblast cells was not due to the difference in surface protein adsorption.Given all the factors,the surfaces obtained by anodic oxidation treatment revealed higher cell viability than those obtained by plasma oxidation(p=0.05).
文摘Although nanocomposites have recently attracted special interest in the tissue engineering area,due to their potential to reinforce scaffolds for hard tissues applications,a number of variables must be set prior to any clinical application.This manuscript addresses the evaluation of thermo-mechanical properties and of cell proliferation of cellulose nanocrystals(CNC),poly(butylene adipate-co-terephthalate)(PBAT),poly(ε-caprolactone)(PCL)films and their bionanocomposites with 2 wt% of CNC obtained by casting technique.Cellulose nanocrystals extracted from Balsa wood by acid hydrolysis were used as a reinforcing phase in PBAT and PCL matrix films.The films and pure CNC at different concentrations were cultured with osteoblasts MG-63 and the cell proliferation was assessed by AlamarBlue?assay.The thermal-mechanical properties of the films were evaluated by dynamic-mechanical thermal analysis(DMTA).It was found by DMTA that the CNC acted as reinforcing agent.The addition of CNCs in the PBAT and PCL matrices induced higher storage moduli due to the reinforcement effects of CNCs.The cell viability results showed that neat CNC favored osteoblast proliferation and both PBAT and PCL films incorporated with CNC were biocompatible and supported cell proliferation along time.The nature of the polymeric matrix or the presence of CNC practically did not affect the cell proliferation,confirming they have no in vitro toxicity.Such features make cellulose nanocrystals a suitable candidate for the reinforcement of biodegradable scaffolds for tissue engineering and biomedical applications.
基金the tobacco cell culture.This work was supported by the n ational n atural Science Foundation of China(n O.31870246,31560059,and 31260059)the Fundamental Research Funds for the Gansu Universities of Gansu Provincial Department of Finance,the University Scientific Research Project of Gansu Province(n o.2015A-007)+1 种基金the Key Research and Development Project of Gansu Province(n o.18YF1 n A051)the Youth Teacher Scientific Research Ability Promotion Plan Innovation Team Project of n orthwest n ormal University.
文摘Cadmium(Cd)is one of the most widespread and toxic heavy metals to plants.Extracellular ATP(exATP)is thought to be an extracellular effector in regulating the physiological responses of plant cells to environmental stresses.However,the function of exATP in Cd-stressed plant cells is much unknown.The present work showed that treating tobacco(Nicotiana tabacum L.cv.Bright Yellow-2)cell-suspension cultures with exogenous CdCl2 reduced the cell viability,exATP level,and Mg content.However,the production of reactive oxygen species(ROS),Cd content,and electrolyte leakage of the cells were enhanced by exogenous CdCl2.When the Cd-induced accumulation of ROS was decreased by the supplement with DMTU(dimethylthiourea,a scavenger of ROS),the Cd-induced increases of the electrolyte leakage and Cd content were alleviated,and the Cd-induced reductions of cell viability were partly rescued,suggesting that Cd-induced reduction of cell viability could be related to the ROS accumulation.Under the condition of Cd stress,when the reduction of exATP level was partly rescued by exogenous ATP(20μM),the increases of ROS production,electrolyte leakage,and Cd content were attenuated,and the reduction of cell viability was also alleviated.These observations indicate that exATP can regulate the cell viability in the Cd–stressed plant cells possibly by an ROS-associated mechanism.
基金Conselho Nacional de Desenvolvimento Cientifico e Tecnologico(CNPq-Brazil)for financial support.
文摘Cellular radiosensitivity is directly correlated with the mechanism of DNA repair, in which p53 protein plays a major role. In this context, this study correlated cell death with p53 expression in lymphocytes irradiated in vitro with different doses of gammaradiation. For this, peripheral blood samples were collected from 10 healthy subjects. Each sample was divided in aliquots and, separately, irradiated with doses of 0,5;2 and 4 Gy. After this, peripheral blood mononuclear cells (PBMCs) were isolated and cultivated during 72 hours in 5% CO2 at 37oC without mitogen stimulation. The expression of p53 protein was evaluated by flow cytometry. In parallel, cell viability was determined by trypan blue staining. Statistical analysis was performed us-ing analysis of variance (ANOVA), differences were considered as statistically significant when p < 0.05. The results showed an increase of p53 expression with the absorbed dose, which was proportional to cell death, suggesting that p53 can be used as bioindicator of individual radiosensitivity.
基金Natural Science Foundation of Shandong Province,China (No.ZR2010HQ041)
文摘·AIM:To transduce recombinant human platelet-derived growth factor B(PDGF-B) gene adeno-associated virus(AAV) to in vitro cultured cat corneal endothelial cell(CEC) and observe the effect of the expressed PDGF-BB protein on the viability of cat CEC.·METHODS:Cat cornea endothelium was torn under microscope and rapidly cultivated in DMEM to form single layer CEC and the passage 2 endothelial cells were used in this study. The recombinant human PDGF-B gene AAV was constructed and transduced into cat CEC directly. Three groups were as following:blank control group, AAV control group and recombinant AAV group. At 24 hours, 48 hours, and 5 days after transduction, total RNA was extracted from the CEC by Trizol and the expression of PDGF-B gene was detected by fluorescence quantitative polymerase chain reaction. Viability of the transduced CEC was detected at 48 hours after transduction by MTT assay. Cell morphology was observed under inverted phase contrast microscope.·RESULTS:With the torn endothelium culture technique, we rapidly got single layer cat CEC. At 24 hours, 48 hours and 5 days after transduction, fluorescence quantitative polymerase chain reaction showed there was no significant difference of the expressed PDGF-B gene mRNA between blank control group and AAV control group (P >0.05). In contrast, there were significant differences between two control groups and recombinant AAV group (P <0.05). MTT assay showed that in recombinant AAV group, the expressed PDGF-BB protein could promote the viability of cat CEC. Morphology observation showed at 48 hours after transduction, cells in CEC-AAV-PDGF-B group proliferated into bigger scales in regular triangle to hexagon shape with distinct boundary, while the number of cells was significantly less in the two control groups.·CONCLUSION:The recombinant AAV-PDGF-B expresses biological active PDGF-BB protein in cat CEC, which promotes the viability and proliferation of cells.
文摘AIM:To investigate the effect of aspirin on neuroendocrine tumor(NET)cell growth and signaling in vitro.METHODS:Human pancreatic BON1,bronchopulmonary NCI-H727 and midgut GOT1 neuroendocrine tumor cells were treated with different concentrations of aspirin(from 0.001 to 5 mmol/L),and the resulting effects on metabolic activity/cell proliferation were measured using cell proliferation assays and SYBR-DNAlabeling after 72,144 and 216 h of incubation.The effects of aspirin on the expression and phosphorylation of several critical proteins that are involved in the most common intracellular growth factor signaling pathways(especially Akt protein kinase B)and mammalian target of rapamycin(mTOR)were determined by Western blot analyses.Propidium iodide staining and flow cytometry were used to evaluate changes in cell cycle distribution and apoptosis.Statistical analysis was performed using a 2-tailed Student’s t-test to evaluate the proliferation assays and cell cycle analyses.The results are expressed as the mean±SD of 3 or 4 independently performed experiments.Statistical significance was set at P<0.05.RESULTS:Treatment with aspirin suppressed the viability/proliferation of BON1,NCI-H727 and GOT1 cells in a time-and dose-dependent manner.Significant effects were observed at starting doses of 0.5-1 mmol/L and peaked at 5 mmol/L.For instance,after treatment with 1 mmol/L aspirin for 144 h,the viability of pancreatic BON1 cells decreased to 66%±13%(P<0.05),the viability of bronchopulmonary NCI-H727 cells decreased to 53%±8%(P<0.01)and the viability of midgut GOT1 cells decreased to 89%±6%(P<0.01).These effects were associated with a decreased entry into the S phase,the induction of the cyclin-dependent kinase inhibitor p21 and reduced expression of cyclindependent kinase 4 and cyclin D3.Aspirin suppressed mTOR downstream signaling,evidenced by the reduced phosphorylation of the mTOR substrates 4E binding protein 1,serine/threonine kinase P70S6K and S6 ribosomal protein and inhibited glycogen synthase kinase3 activity.We observed the(compensatory)activation of tuberous sclerosis 2,the serine/threonine specific protein kinase AKT and extracellular signal-regulated kinases.CONCLUSION:Aspirin demonstrates promising anticancer properties for NETs in vitro.Further preclinical and clinical studies are needed.
基金supported partly by National Natural Science Foundation of China(Nos.81372076,51307133 and 51221005)China National Funds for Distinguished Young Scientists(No.51125029)+1 种基金the Sci-Tech Project of Shaanxi Province of China(No.2010K16-04)the Fundamental Research Funds for the Central Universities of China(No.xkjc2013004)
文摘An argon atmospheric pressure plasma jet was employed to treat L929 murine fibroblasts cultured in vitro.Experimental results showed that,compared with the control cells,the treatment of fibroblasts with 15 s of plasma led to a significant increase of cell viability and collagen synthesis,while the treatment of 25 s plasma resulted in a remarkable decrease.Exploration of related mechanisms suggested that cold plasma could up-regulate Cyclin D1 gene expression and down-regulate p27 gene expression at a low dose,while it could down-regulate Cyclin D1 expression and up-regulate p27 expression at a higher dose,thus altering the cell cycle progression,and then affecting cell viability and collagen synthesis of fibroblasts.
文摘AIM: To analyze the concentration-dependent effects of autologous serum(AS) and fetal bovine serum(FBS) on human corneal epithelial cell(HCEC) viability, migration and proliferation.METHODS: AS was prepared from 13 patients with nonhealing epithelial defects Dulbecco's modified eagle medium/Ham's F12(DMEM/F12) with 5% FBS, 0.5% dimethyl sulphoxide(DMSO), 10 ng/m L human epidermal growth factor, 1% insulin-transferrin-selenium, then were incubated in serum media: DMEM/F12 supplemented by 5%, 10%, 15% or 30% AS or FBS. HCEC viability was analyzed using cell proliferation kit XTT, migration using a wound healing assay, proliferation by the cell proliferation enzyme-linked immunosorbent assay(ELISA) Brd U kit. Statistical analysis was performed using the generalized linear model, the values at 30% AS or 30% FBS were used as the baselines.RESULTS: HCEC viability was the highest at 30% AS or 15% FBS and the lowest at 10% AS or 30% FBS application. HCEC migration was the quickest through 30% AS or 30% FBS and the slowest through 5% AS or 5% FBS concentrations. Proliferation was the most increased through 15% AS or 5% FBS and the least increased through 30% AS or 30% FBS concentrations. HCEC viability at 10% and 15% AS was significantly worse(P=0.001, P=0.023) compared to baseline and significantly better at 15% FBS(P=0.003) concentrations. HCEC migration was significantly worse(P≤0.007) and HCEC proliferation significantly better(P<0.001) in all concentration groups compared to baseline.CONCLUSION: For the best viability of HCEC 30% AS or 15% FBS, for HCEC migration 30% AS or 30% FBS, for proliferation 15% AS or 5% FBS should be used. Therefore, we suggest the use of 30% AS in clinical practice.
文摘Background: Mammalian ovaries contain follicles containing an oocyte enclosed by layers of granulosa cells (GC). Follicle growth and oocyte maturation are largely dependent on GC numbers and viability, but there is no established, reliable method for assessing the number of viable GC within an isolated follicle. Methods: Centrifugation conditions and the Trypan Blue (TB) Exclusion assay were optimised for low cell densities compatible with the numbers of GC in follicles. Mouse ovarian follicles were disaggregated to produce a single cell suspension of GC which were examined by TB (n = 4), but also by crystal violet assay in a 96-well plate format after 24 h in vitro (n = 3). GC viability in vitro was characterised further by using enzyme-linked immunoassays to quantify GC production of anti-Mullerian hormone (AMH) and estrogen. Results: The centrifugation and low cell density TB protocol could accurately measure the viability of 78 GC in 10 μL, with an intra-assay coefficient of variation (CoV) 22%, and inter-assay CoV 7%. The best follicle disaggregation method (30 min 37°C exposure to 2 mg/mL collagenase prior to 30 min exposure to 0.025% hyaluronidase) yielded (656 ±87) GC per antral follicle of which 82% ±5% were viable. Culturing 312 - 20,000 GC per well for 24 hours and assessing viability by crystal violet assay generated a linear correlation between OD value and viable GC number (R2 = 0.98) and estrogen concentration per well (R2 = 0.92). 20,000 GC per well produced 143 ±16 pg/mL estrogen during 24 hours in vitro, but no detectable AMH. Conclusion: This is the first report describing the isolation of viable, estrogen-producing GC from murine follicles, and their subsequent culture. These procedures are transferrable to other species including humans and can be applied to screening the reproductive toxicity of pharmaceutical agents.
文摘Background: Despite the popularity of autologous fat transfer applications, high resorption rates, and consequential volume loss, have been reported. Viable adipocyte content has been defined as a key determinant of fat transfer longevity. Moreover, traces of blood, free oil fat and fibrotic tissue accelerate adipocyte degradation. Objective: To compare the effectiveness of a 1470 nm, radial emitting laser-assisted liposection device to a mechanical liposection device in maintaining adipocyte viability in fat tissue harvests. Methods: Bilateral subcutaneous adipose tissue samples were harvested from ten female patients. Fat was harvested from one side using the LipoLife laser-assisted liposuction device and from the other side with a Byron mechanical aspirator. Samples were visually analyzed and blood:fat ratios and cell viability were determined. Results: Laser-harvested samples separated into two distinct phases, with a negligible blood phase at the bottom (1.1%) and a significant adipose phase at the top (98.9%), containing small, uniform-sized cells, of which 95.7% ± 2.7% proved viable. Mechanically harvested samples separated into blood (18%), adipose (60%) and lipid (22%) phases. The adipose phase contained significant amounts of connective tissue, large adipose tissue fragments, large oil droplets and a mean 79.7% ± 18.3% viable adipocytes. Conclusions: Laser liposuctioning was superior to mechanical liposuctioning, providing both higher cell viability and enhanced sample quality. The 1470 nm diode laser bears the potential of improving long-term clinical outcomes of fat transfer procedures. Improved purity of the harvested sample and heightened preadipocyte content are projected to provide for extended graft longevity.
文摘Vaccination with tumor-antigen pulsed, monocyte-derived dendritic cells (DCs) has emerged as a promising strategy in cancer immunotherapy. The standard DC maturation cocktail consists of a combination of tumor necrosis factor-α (TNF-α)/interleukin (IL)-1β/IL-6 and prostaglandin E2 (PGE2) for generation of standard DCs (sDCs). In order to improve IL-12p70 production and cytotoxic T-lymphocyte (CTL) induction, a novel cocktail composed of TNF-α/IL-1β/ interferon (IFN)-α/IFN-γ and polyinosinic:polycytidylic acid (Poly-I:C) has been introduced to generate so-called α-Type-1 polarized DCs (αDC1s). We and others have previously performed a comprehensive comparison of sDCs and αDC1s. Here we demonstrate that the viability of αDC1s is lowered compared to sDCs and that DC apoptosis is mediated by Poly-I:C. We speculated that activation of protein kinase R (PKR) could mediate the observed apoptosis, but despite significantly higher PKR expression in αDC1s compared to sDCs and induction of active threonine (Thr)446 autophosphorylation of PKR in αDC1s, Poly-I:C did not influence total PKR expression or autophosporylation, indicating PKR-independent Poly-I:C-induced DC apoptosis.
基金supported by the Science and Technology Development Foundation of Beijing Science and Technology Commission, No. Z101107052210004
文摘In the present study, PC12 cells induced by 6-hydroxydopamine as a model of Parkinson's Disease, were used to investigate the protective effects of bone marrow-derived mesenchymal stem cells bone marrow-derived mesenchymal stem cells against 6-hydroxydopamine-induced neurotoxicity and to verify whether the mechanism of action relates to abnormal α-synuclein accumulation in cells. Results showed that co-culture with bone marrow-derived mesenchymal stem cells enhanced PC12 cell viability and dopamine secretion in a cell dose-dependent manner. MitoLight staining was used to confirm that PC12 cells co-cultured with bone marrow-derived mesenchymal stem cells demonstrate reduced levels of cell apoptosis. Immunocytochemistry and western blot analysis found the quantity of α-synuclein accumulation was significantly reduced in PC12 cell and bone marrow-derived mesenchymal stem cell co-cultures. These results indicate that bone marrow-derived mesenchymal stem cells can attenuate 6-hydroxydopamine-induced cytotoxicity by reducing abnormal α-synuclein accumulation in PC12 cells.
文摘It is well known that α-synuclein (αS) plays an important role in the pathogenesis of Parkinson’s disease (PD). Moreover, oxidative stress is also thought to be an important factor in PD due to induction of dopaminergic neuronal cell death by free radicals and enhancement of αS fibrillation by oxidized stress. In the present study, to clarify the role of glutathione (GSH), an intracellular antioxidant, on the molecular mechanism of αS-induced cell injury, we examined the effects of L-buthionine-SR-sulfoximine (BSO), a GSH synthase inhibitor, with or without N-acetyl-L-cysteine (NAC), a source of GSH, on αS-induced cell injury in human neuroblastoma SH-SY5Y cells. Treatment with BSO significantly reduced the cell viability of both empty-vector- and αS-transfected SH-SY5Y cells in a dose-dependent manner (p < 0.01), although the ratio of αS-induced reduction of cell viability in α-syn-transfected cells was much greater than that in empty-vector-transfected cells. Moreover, BSO significantly reduced the intracellular total GSH level in both types of transformant cells. However, NAC significantly prevented BSO-induced reduction of both cell viability and GSH level in the αS-transfected cells. These findings suggest that GSH plays an important role in αS-induced cell injury by reducing cell viability.
基金supported by National Natural Science Foundation of China (Nos. 30570442, 10225526)Hundred Talents Program of The Chinese Academy of Sciences and Foundation of President, of The Hefei Institutes of Physical Sciences, CAS
文摘Magnetic resonance image (MRI) systems with a much higher magnetic flux densitywere developed and applied for potential use in medical diagnostic. Recently, much attention hasbeen paid to the biological effects of static, strong magnetic fields (SMF). With the 13 T SMFfacility in the Institute of Plasma Physics, Chinese Academy of Sciences, the present study focusedon the cellular effects of the SMF with 13 T on the cell viability and the cell cycle distributionin immortalized hamster cells, such as human-hamster hybrid (A_L) cells, Chinese hamster ovary(CHO) cells, DNA double-strand break repair deficient mutant (XRS-5) cells, and human primaryskin fibroblasts (AG1522) cells. It was found that the exposure of 13 T SMF had less effect onthe colony formation in either nonsynchronized or synchronized A_L cells. Moreover, as comparedto non-exposed groups, there were slight differences in the cell cycle distribution no matter ineither synchronized or nonsynchronized immortalized hamster cells after exposure to 13 T SMF.However, it should be noted that the percentage of exposed AG1522 cells at G0/G1 phase wasdecreased by 10% as compared to the controls. Our data indicated that although 13 T SMF hadminimal effects in immortalized hamster cells, the cell cycle distribution was slightly modified bySMF in human primary fibroblasts.
文摘The traditional MTT assay requires destructive analyses and is not convenient in continuous monitory of cell viability. However, a new cell model was developed in this research, by using the oxidation-reduction (redox) indicator alamarblue? instead of the MTT assay. The alamarblue?does no harm to cells and provide a much more safe and convenient methods of measurement. Firstly, cell apoptosis was induced by different concentration of 6-OHDA, then the cell viability was tested by the alamarblue?at a serial of time points. Finally, the optimism cells density, 6-OHDA concentration and testing time point were gained to set a wonderful SH-SY5Y cell model in our research. And when it is applied in the study of neuroprotection effects of NAC, GSH and Catalase, the new model reveals undeniable advantages.
基金Supported by Important Subject Fund of Science Technology Department of Zhejiang Province(No.2013C03048-1)
文摘AIM:To investigate the impact of adipose-derived mesenchymal stem cells(ADSCs) on cell viability and extracellular matrix(ECM) synthesis of corneal stromal cells(CSCs). METHODS:ADSCs and CSCs were obtained from the corneas of New Zealand white rabbits and indirectly cocultured in vitro. The proliferative capacity of CSCs in the different groups was assessed by CCK-8 assays. Annexin V-fluorescein isothiocyanate(FITC)/proliferation indices(PI) assays were used to detect the apoptosis of CSCs. The expression levels of matrix metalloproteinase(MMP), such as MMP1, MMP2, MMP9, and collagens were also evaluated by Western blot. RESULTS:ADSCs significantly promoted proliferation and invasion of CSCs in the indirect co-culture assays. The co-cultural group displayed much higher ability of proliferation, especially under the co-culture conditions of ADSCs for 3d, compared with that CSCs cultured alone. The PI of CSCs in the co-culture system were increased approximately 3-8-fold compared with the control group. A significant change was observed in the proportions of cells at apoptosis(early and late) between the negative control group(6.34% and 2.06%) and the ADCSs-treated group(4.69% and 1.59%). The expression levels of MMPs were down regulated in the co-culture models. Compared with the control group, the decrease intensities of MMP-1, MMP-2 and MMP-9 in CSCs/ADSCs group were observed, 3.90-fold, 1.09-fold and 3.03-fold, respectively. However, the increase intensities of collagen type(I, II, III, IV, and V) in CSCs were observed in CSCs/ADSCs group, 3.47-fold,4.30-fold, 2.35-fold, 2.55-fold and 2.43-fold, respectively, compared to that in the control group. The expressions of aldehyde dehydrogenase and fibronectin in CSCs were upregulated in the co-culture models.CONCLUSION:ADSCs play a promotive role in CSCs' growth and invasion, which may be partially associated with MMPs decrease and collagens increase, resulting in a positive participation in the plasticity and ECM synthesis of CSCs. This provided a new insight into the extensive role of ADSCs in CSCs and a potential molecular target for corneal therapy.
