Neuroprotective effects of genistein on SH-SY5Y cells overexpressing A53T mutant α-synuclein

Genistein, a potent antioxidant compound, protects dopaminergic neurons in a mouse model of Parkinson＇s disease. However, the mecha- nism underlying this action remains unknown. This study investigated human SH-SYSY cells overexpressing the A53T mutant of α-synuclein. Four groups of cells were assayed： a control group （without any treatment）, a genistein group （incubated with 20 μM genistein）, a rote- none group （treated with 50 μM rotenone）, and a rotenone ＋ genistein group （incubated with 20 μM genistein and then treated with 50μM rotenone）. A lactate dehydrogenase release test confirmed the protective effect of genistein, and genistein remarkably reversed mitochondrial oxidative injury caused by rotenone. Western blot assays showed that BCL-2 and Beclin ! levels were markedly higher in the genistein group than in the rotenone group. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling revealed that genistein inhibited rotenone-induced apoptosis in SH-SYSY cells. Compared with the control group, the expression of NFE2L2 and HMOX1 was significantly increased in the genistein ＋ rotenone group. However, after treatment with estrogen receptor and NFE2L2 channel blockers （ICI-182780 and ML385, respectively）, genistein could not elevate NFE2L2 and HMOX1 expression. ICI-182780 effectively prevented genistein-mediated phosphorylation of NFE2L2 and remarkably suppressed phosphorylation of AKT, a protein downstream of the estrogen receptor. These findings confirm that genistein has neuroprotective effects in a cell model of Parkinson＇s dis- ease. Genistein can reduce oxidative stress damage and cell apoptosis by activating estrogen receptors and NFE2L2 channels.

Neuroprotective effects of bloodletting at Jing points combined with mild induced hypothermia in acute severe traumatic brain injury

Bloodletting at Jing points has been used to treat coma in traditional Chinese medicine. Mild induced hypothermia has also been shown to have neuroprotective effects. However, the therapeutic effects of bloodletting at Jing points and mild induced hypothermia alone are limited. Therefore, we investigated whether combined treatment might have clinical effectiveness for the treatment of acute severe traumatic brain injury. Using a rat model of traumatic brain injury, combined treatment substantially alleviated cerebral edema and bloodbrain barrier dysfunction. Furthermore, neurological function was ameliorated, and cellular necrosis and the inflammatory response were lessened. These findings suggest that the combined effects of bloodletting at Jing points（20 μL, twice a day, for 2 days） and mild induced hypothermia（6 hours） are better than their individual effects alone. Their combined application may have marked neuroprotective effects in the clinical treatment of acute severe traumatic brain injury.

Underlying mechanism of protection from hypoxic injury seen with n-butanol extract of Potentilla anserine L. in hippocampal neurons

The alcohol and n-butanol extract of Potentilla anserine L. significantly protects myocardium from acute ischemic injury. However, its effects on rat hippocampal neurons and the mechanism of protection remain unclear. In this study, primary cultured hippocampal neurons from neonatal rats were incubated in 95% N2 and 5% CO2 for 4 hours. Results indicated that hypoxic injury decreased the viability of neurons, increased the expression levels of caspase-9 and caspase-3 mRNA, as well as cytochrome c, Caspase-9, and Caspase-3 protein. Pretreatment with 0.25, 0.062 5, 0.015 6 mg/mL n-butanol extract of Potentilla anserine L. led to a significant increase in cell viability. Expression levels of caspase-9 and caspase-3 mRNA, as well as cytochrome c, Caspase-9, and Caspase-3 protein, were attenuated. The neuroprotective effect of n-butanol extract of Potentilla anserine L. was equivalent to tanshinone IIA. Our data suggest that the n-butanol extract of Potentilla anserine L. could protect primary hippocampal neurons from hypoxic injury by deactivating mitochondrial cell death.

Mitochondrial redox metabolism in aging:Effect of exercise interventions

Mitochondrial redox metabolism has long been recognized as being central to the effects of aging and the development of age-related pathologies in the major oxidative organs. Consistent evidence has shown that exercise is able to retard the onset and impede the progression of aging by modifying mitochondrial oxidant--antioxidant homeostasis. Here we provide a broad overview of the research evidence showing the relationship between mitochondrial redox metabolism, aging and exercise. We address part aspects of mitochondrial reactive oxygen species （ROS） metabolism, from superoxide production to ROS detoxification, especially antioxidant enzymes and uncoupling protein. Furthermore, we describe mitochondrial remodeling response to aging and exercise, which is accompanied by bioenergetics and redox regulation. In addition, potential mechanisms for redox signaling involved in mitochondrial remodeling and redox metabolism regulation are also reviewed.

Double laminated reduced graphene/Cu_2S/reduced graphene/graphene oxide nanofilms and their photoelectrochemical properties

In this work, an efficient photocatalytic material was prepared directly on Indium tin oxide (ITO) glass substrates by fabricating Cu2S and graphene oxide onto graphene for photoelectrochemical (PEC) water splitting. The double laminated reduced graphene/Cu2S/reduced graphene/graphene oxide (RG/Cu2S/RG/GO) nanofilms were characterized, and an enhanced photoelectrochemical response in the visible region was discovered. The photocurrent density of the nanofilms for PEC water splitting was measured to be up to 1.98 mA/cm(2), which could be ascribed to the followings: (i) a higher efficiency of light-harvesting because of GO coupling with Cu2S that could broaden the absorbing solar spectrum and enhance the light utilization efficiency; (ii) a stepwise structure of band-edge levels in the Cu2S/GO electrode was constructed; (iii) double laminated electron accelerator (RG) was used in the Cu2S/GO materials to get better electron-injecting efficiency. (C) 2016 Science Press and Dalian Institute of Chemical Physics, Chinese Academy of Sciences. Published by Elsevier B.V. and Science Press. All rights reserved.

Clinical efficacy of radiofrequency ablation for primary hepatocellular carcinoma

Objective:To summarize and analyze the effects of radiofrequency ablation (RFA) on patients' condition and quality of life after treatment of large liver cancer.Methods: A total of 51 patients with primary liver cancer with a diameter of 4-7 cm were selected from our hospital. The related tumor markers and liver function indexes were analyzed before and after treatment, and the patients' progression-free survival was analyzed with long-term follow-up.Results: On the 3rd day after the treatment, the AFP level of the patients was not significantly different from that before the treatment (P>0.05), and on the 7th day after the treatment, the AFP level was significantly lower than that before the treatment (P<0.05). The levels of CA199 and ferritin were slightly higher on the 3rd day after treatment than before treatment (P<0.05), and the levels of CA199 and ferritin were significantly lower on the 7th day after treatment than before treatment (P<0.05). The index of liver function at 3 days after treatment was slightly higher than that before treatment (P<0.05), and there was no significant difference in liver function at 7 d after treatment (P>0.05). Progression-free survival was observed at 1 month (90.2%), 38 at 3 months (74.5%), 33 at 6 months (64.7%), and 28 at 12 months (54.9%). After 1 month of treatment, the ECOG of 48 patients recovered to the pre-treatment level, with 3 cases of deterioration (including 2 deaths);After 3 months of treatment, there was no significant change in ECOG in 42 patients, and the ECOG score deteriorated in 9 patients (including 3 deaths). After 6 months of treatment, 36 cases of ECOG were unchanged, while 12 cases were deteriorated (including 5 deaths). After 12 months of treatment, 30 cases of ECOG were unchanged, with 21 cases of deterioration (including 9 deaths).Conclusion: Radiofrequency ablation can better alleviate the disease and improve the quality of life of patients with primary liver cancer with diameter of 4-7 cm. Further studies are needed to compare the effects of other local treatments.

Biomedical response of femurs in male Wistar rat in chronic hypergravity environments

Bone is sensitive to mechanical stimulation and plays a loading-bearing role in the human body.However,regulation of bone biomechanical properties in chronic hypergravity environments is still unclear.In this study,male Wistar rats exposed to chronic hypergravity environments(4
g,8
g,10
g,and 20
g)for 4 weeks were set as the hypergravity groups,and rats exposed to the normal gravity as the control group.Morphology parameters and bone remodeling factors were obtained by means of micro-CT,Western blot,and q-PCR.Mechanical properties of femurs were measured utilizing three points bending test and creep test and were fitted into a viscoelastic-viscoplastic constitutive equation.The results indicate osteoporosis occurred in femurs of hypergravity groups.Accordingly,the protein and gene expressions of bone remodeling factors(OPG,RANKL,runx2)in hypergravity groups were significantly different from that in the control group,demonstrating that bone formation level increased and bone resorption level decreased.Meanwhile,mechanical properties of femurs in hypergravity groups showed that Young's modulus of femurs in the 20
g group was significantly higher than that in the control group.The viscoelastic-viscoplastic properties of bone tissue were changed in hypergravity environments.Among them,the 8
g group was closest to the control group in morphology and mechanical properties.To sum up,the biomechanical response regulation of rat femur under 4-20
g chronic hypergravity environments was presented.Hypergravity environments could lead to osteoporosis.The balance between bone formation and bone resorption would be disrupted in hypergravity groups due to bone adaptation.20
g environment has a significant effect on elastic modulus on femurs.Due to the difference in biomechanical response of femurs,the viscoelastic-viscoplastic characteristics of femurs have a nonlinear relationship with hypergravity values.Bone tissue was least affected by 8
g hypergravity in morphology and mechanical properties.

Network toxicology and LC-MS-based metabolomics:New approaches for mechanism of action of toxic components in traditional Chinese medicines

Network toxicology combined with metabonomics is of great significance for the study of the toxic mechanism and prediction of toxicity of traditional Chinese medicines(TCMs).In this study,we reviewed the application of network toxicology based on LC-MS metabolomics,mainly in the study of toxic components and the toxicity mechanism of TCMs,which provides new ideas and methods for the further study of the toxicity mechanism of TCMs.

Saikosaponin a increases interleukin-10 expression and inhibits scar formation after sciatic nerve injury

Nerve scarring after peripheral nerve injury can severely hamper nerve regeneration and functional recovery.Further,the anti-inflammatory cytokine,interleukin-10,can inhibit nerve scar formation.Saikosaponin a（SSa） is a monomer molecule extracted from the Chinese medicine,Bupleurum.SSa can exert anti-inflammatory effects in spinal cord injury and traumatic brain injury.However,it has not been shown whether SSa can play a role in peripheral nerve injury.In this study,rats were randomly assigned to three groups.In the sham group,the left sciatic nerve was directly sutured after exposure.In the sciatic nerve injury（SNI） ＋ SSa and SNI groups,the left sciatic nerve was sutured and continuously injected daily with SSa（10 mg/kg） or an equivalent volume of saline for 7 days.Enzyme linked immunosorbent assay results demonstrated that at 7 days after injury,interleukin-10 level was considerably higher in the SNI ＋ SSa group than in the SNI group.Masson staining and western blot assay demonstrated that at 8 weeks after injury,type I and III collagen content was lower and nerve scar formation was visibly less in the SNI ＋ SSa group compared with the SNI group.Simultaneously,sciatic functional index and nerve conduction velocity were improved in the SNI ＋ SSa group compared with the SNI group.These results confirm that SSa can increase the expression of the anti-inflammatory factor,interleukin-10,and reduce nerve scar formation to promote functional recovery of injured sciatic nerve.

ZM-66, a New Podophyllotoxin Derivative Inhibits Proliferation and Induces Apoptosis in K562/ADM Cells

Objective To investigate the anti-tumor effect of ZM-66 on multidrug-resistant leukemic cell line K562/ADM. Methods The K562/ADM cells were treated with varying concentrations （0, 1, 2, 4×10^-3 mmol/L） of ZM-66 or etoposide for 24 hours. The proliferation was detected by Sulforhodamine B Sodium Salt （SRB） assay and apoptosis was detected by flow cytometry analysis and fluorescent staining. In addition, the expression levels of p53 and bax genes in K562/ADM cells were detected by RT-PCR analysis. The level of P-glycoprotein （P-gp）, P53 and Bax protein in K562/ADM cells were detected by Western blot assay. Results SRB assay demonstrated that etoposide had little inhibitory effect on K562/ADM cells, whereas ZM-66 （1, 2, 4×10^-3 mmol/L） had significantly inhibitory effect on K562/ADM cells （all P〈0.01）. The acridine orange/propidium iodide dual staining showed that there were typical condensation of chromatin and nuclear fragmentation nuclei with red color in ZM-66 treated cells. Flow cytometric analysis showed that there was a significantly increase of apoptotic cells i~ K562/KDM cells after treated with ZM-66. RT-PCR showed that the p53 and bax mRNA expression levels in K562/ADM cells treated with ZM-66 at 1, 2, 4×10^-3 mmol/L were higher than those in the cell without treatment. Western blot showed that the P53 and Bax protein expression levels in K562/ADM cells treated with ZM-66 at 2, 4x 10 s mmol/L were higher than those in the cell without treatment. But the P-gp protein expression level in K562/ADM cells treated with ZM-66 at 2, 4×10^-3 mmol/L was gradually lower than those in the cell without treatment. Conclusion ZM-66 is able to induce cell death by apoptosis in vitro, as a result of the reverse of theapoptosis resistance in drug-resistant K562/ADM cells by modulating expression of key factors associated with apoptosis induction.

Magnetic resonance imaging-three-dimensional printing technology fabricates customized scaffolds for brain tissue engineering

Conventional fabrication methods lack the ability to control both macro- and micro-structures of generated scaffolds. Three-dimensional printing is a solid free-form fabrication method that provides novel ways to create customized scaffolds with high precision and accuracy. In this study, an electrically controlled cortical impactor was used to induce randomized brain tissue defects. The overall shape of scaffolds was designed using rat-specific anatomical data obtained from magnetic resonance imaging, and the internal structure was created by computer- aided design. As the result of limitations arising from insufficient resolution of the manufacturing process, we magnified the size of the cavity model prototype five-fold to successfully fabricate customized collagen-chitosan scaffolds using three-dimensional printing. Results demonstrated that scaffolds have three-dimensional porous structures, high porosity, highly specific surface areas, pore connectivity and good internal characteristics. Neural stem cells co-cultured with scaffolds showed good viability, indicating good biocompatibility and biodegradability. This technique may be a promising new strategy for regenerating complex damaged brain tissues, and helps pave the way toward personalized medicine.

Exendin-4 inhibits high-altitude cerebral edema by protecting against neurobiological dysfunction

The anti-inflammatory and antioxidant effects of exendin-4（Ex-4） have been reported previously.However,whether（Ex-4） has anti-inflammatory and antioxidant effects on high-altitude cerebral edema（HACE） remains poorly understood.In this study,two rat models of HACE were established by placing rats in a hypoxic environment with a simulated altitude of either 6000-or 7000-m above sea level（MASL） for 72 hours.An altitude of 7000 MASL with 72-hours of hypoxia was found to be the optimized experimental paradigm for establishing HACE models.Then,in rats where a model of HACE was established by introducing them to a 7000 MASL environment with 72-hours of hypoxia treatment,2,10 and,100 μg of Ex-4 was intraperitoneally administrated.The open field test and tail suspension test were used to test animal behavior.Routine methods were used to detect change in inflammatory cells.Hematoxylin-eosin staining was performed to determine pathological changes to brain tissue.Wet/dry weight ratios were used to measure brain water content.Evans blue leakage was used to determine blood-brain barrier integrity.Enzyme-linked immunosorbent assay（ELISA） was performed to measure markers of inflammation and oxidative stress including superoxide dismutase,glutathione,and malonaldehyde values,as well as interleukin-6,tumor necrosis factor-alpha,cyclic adenosine monophosphate levels in the brain tissue.Western blot analysis was performed to determine the levels of occludin,ZO-1,SOCS-3,vascular endothelial growth factor,EPAC1,nuclear factor-kappa B,and aquaporin-4.Our results demonstrate that Ex-4 preconditioning decreased brain water content,inhibited inflammation and oxidative stress,alleviated brain tissue injury,maintain blood-brain barrier integrity,and effectively improved motor function in rat models of HACE.These findings suggest that Ex-4 exhibits therapeutic potential in the treatment of HACE.

Saikosaponin A induces apoptosis and upregulates autophagy in Huh7 cells

Objective: It is discussed whether saikosaponin A induces apoptosis of human hepatoma Huh7 cells is related to the change of autophagy level.Methods: The effects of different concentrations of SSA on proliferation and apoptosis of Huh7 cells were detected by MTT and flow cytometry, and then constructed recombinant plasmid pEGFP-N1-LC3B and transfected into Huh7 cells. After intervened by SSA culture medium, the autophagy level was observed under confocal microscope. The expression of apoptosis proteins Bax, Bcl-2, PCNA and autophagy-related proteins LC3B, Beclin1, and Apg12-Apg5 were detected by Western Blot. Results: SSA can significantly inhibit the proliferation of Huh7 cells, promote apoptosis, increase the number of autophagy bodies in the cytoplasm, up-regulate the expression of Bax, LC3B-II, Beclin1, Apg12-Apg5 and down-regulate the expression of Bcl-2, PCNA. Conclusion:SSA induced apoptosis of Huh7 cells in vitro and upregulated the autophagy level.

Neuroprotection of n-Butanol Extract from Roots of Potentilla anserina on Hypoxic Injury in Primary Hippocampal Neurons

Objective To investigate the protective effect of n-butanol extract from the roots of Potentilla anserina (NP) on hypoxic hippocampal neurons in neonatal rats. Methods Primary cultured hippocampal neurons were pretreated with different concentration of NP (0.25, 0.0625, and 0.0156 mg/mL) before incubation in a low oxygen (0.1%) environment for 4 h. Cell viability was evaluated by Trypan blue staining assay. Lactate dehydrogenase (LDH) released by neurons into the medium was measured. The activity of superoxide dismutase (SOD) in cell cytosol was determined using nitroblue tetrazolium. Morphological changes and mitochondrial function were observed by transmission electron microscopy. Results Hypoxic injury could decrease the cells viability of neuron, enhance LDH release (P < 0.05), decrease SOD activity, and increase mitochondrial injury. Pretreatment with NP significantly increased cell viability, decreased LDH release (P < 0.05), promoted SOD activity (P < 0.05), and remarkably improved cellular ultra-microstructure compared with the model group. Conclusion NP could protect the primary hippocampal neurons from hypoxic injury by attenuating mitochondrial cell death.

Effect of n-butanol Extract from Potentilla anserina on Hypoxia-induced Calcium Overload and SERCA2 Expression of Rat Cardiomyocytes

Objective To investigate the effect of n-butanol extract from Potentilla anserina(NP)intervention on hypoxia-induced Ca 2+ overload and SERCA2 expression of rat cardiomyocytes.Methods Primary cultured myocardial cell from SD neonatal rat(1-3 d)was used in the establishment of hypoxia model.After hypoxia for 3 h,the Ca 2+ concentration of myocardial cells was measured with fura-2/AM fluorescent probe,and the biochemical indicator intracellular Ca 2+ -ATPase was examined and the mRNA and its protective protein levels of the sarcoplasmic reticulum(SR)Ca 2+ -ATPases(SERCA2)were assayed with RT-PCR,Western-blotting,and immune-cytochemical staining in each group.Results The results showed that NP decreased Ca 2+ concentration, increased the activity of Ca 2+ -ATPase,and improved the mRNA and protein expression of SERCA2 in hypoxia-injured myocardial cells as compared with the model group.Conclusion These results indicate that NP could attenuate the Ca 2+ overload.The mechanism might be explained as that NP could elevate the SERCA2 level, increase the activity of myocardium in rats,and further enhance the capacity of SR Ca 2+ re-uptake.

Synthesis and anti-tumor activity evaluation of novel podophyllotoxin derivatives

In order to investigate the effects on the cytotoxicity of indole-3-oxalylamino podophyllotoxin analogs, seven novel podophyllotoxin derivatives were synthesized. The compounds were tested against Hela, K562 or K562/A02 cancer cells in vitro, four of which showed significant cytotoxicity. Among them 9a, 9b and 9c were superior to the positive control VP-16.

Influence of S3 electrical stimulation on gastrointestinal dysfunction after spinal cord injury in rabbits

Objective：To investigate the effect of electrical stimulation to sacral spinal nerve 3 （S3 stimulation） on gastrointestinal dysfunction after spinal cord injury （SCI）.Methods：Six rabbits were taken as normal controls to record their gastrointestinal multipoint biological discharge,colon pressure and rectoanal inhibitory reflex.Electrodes were implanted into S3 in another 18 rabbits.Then the model of SCI was conducted following Fehling＇s method：the rabbit S3 was clamped to induce transverse injury,which was claimed by both somatosensory evoked potential and motion evoked potential.Two hours after SCI,S3 stimulation was conducted.The 18 rabbits were subdivided into 3 groups to respectively record their gastrointestinal electric activities （n=6）,colon pressure （n=6）,and rectum pressure （n=6）.Firstly the wave frequency was fixed at 15 Hz and pulse width at 400 μs and three stimulus intensities （6 V,8 V,10 V） were tested.Then the voltage was fixed at 6 V and the pulse width changed from 200 μs,400 μs to 600 μs.The response was recorded and analyzed.The condition of defecation was also investigated.Results：After SCI,the mainly demonstrated change was dyskinesia of the single haustrum and distal colon.The rectoanal inhibitory reflex almost disappeared.S3 stimulation partly recovered the intestinal movement after denervation,promoting defecation.The proper stimulus parameters were 15 Hz,400 μs,6 V,10 s with 20 s intervals and 10 min with 10 min intervals,total 2 h.Conclusion：S3 stimulation is able to restore the intestinal movement after denervation （especially single haustrum and distal colon）,which promotes defecation.

In vivo skin penetration and metabolic path of quantum dots

The skin is the largest organ of the body and is a potential route of exposure to sunscreens and cosmetics containing nanoparticles; however, the permeability of the skin to these nanoparticles is currently unknown. In this paper, we studied the transderreal delivery capacity through mouse skin of water-soluble CdSeS quantum dots （QDs） and the deposition of these QDs in the body. QD solution was coated onto the dorsal hairless skin of male ICR mice. Fluorescence microscopy and transmission electron microscopy （TEM） were used to observe the distribution of QDs in the skin and organs, and inductively coupled plasma-mass spectrometry （ICP-MS） was used to measure the 111Cd content to indicate the concentration of QDs in plasma and organs. Experimental results indicate that QDs can penetrate into the dermal layer and are limited to the uppermost stratum corneum layers and the hair follicles. Through blood circulation, QDs deposit mostly in liver and kidney and are difficult to clear, 111Cd concentration was greater than 14 ng g-1 in kidney after 120 h after 0.32 nmol QDs was applied to a mouse. These results suggest that QDs have in vivo transdermal delivery capacity through mouse skin and are harmful to the liver and kidney.