Role of XIAP gene overexpressed bone marrow mesenchymal stem cells in the treatment of cerebral injury in rats with cerebral palsy.

BACKGROUND: This study is performed to investigate the effects of adenovirus-mediated X-linked inhibitor of apoptosis protein (XIAP) overexpressed bone marrow mesenchymal stem cells (BMSCs) on brain injury in rats with cerebral palsy (CP). METHODS: Rat's BMSCs were cultured and identified. The XIAP gene of BMSCs was modified by adenovirus expression vector Ad-XIAP-GFP. The rat model of CP with ischemia and anoxia was established by ligating the left common carotid artery and anoxia for 2 h, and BMSCs were intracerebroventricularly injected to the modeled rats. The mRNA and protein expression of XIAP in brain tissue of rats in each group was detected by RT-qPCR and western blot analysis. The neurobehavioral situation, content of acetylcholine (Ach), activity of acetylcholinesterase (AchE), brain pathological injury, apoptosis of brain nerve cells and the activation of astrocytes in CP rats were determined via a series of assays. RESULTS: Rats with CP exhibited obvious abnormalities, increased Ach content, decreased AchE activity, obvious pathological damage, increased brain nerve cell apoptosis, as well as elevated activation of astrocyte. XIAP overexpressed BMSCs improved the neurobehavioral situation, decreased Ach content and increased AchE activity, attenuated brain pathological injury, inhibited apoptosis of brain nerve cells and the activation of astrocytes in CP rats. CONCLUSION: Our study demonstrates that XIAP overexpressed BMSCs can inhibit the apoptosis of brain nerve cells and the activation of astrocytes, increase AchE activity, and inhibit Ach content, so as to lower the CP caused by cerebral ischemia and hypoxia in rats.

Inhibition of Calcium/Calmodulin-Dependent Protein Kinase IIα Suppresses Oxidative Stress in Cerebral Ischemic Rats Through Targeting Glucose 6-Phosphate Dehydrogenase.

Ischemic stroke is a leading cause of mortality and morbidity worldwide, and oxidative stress plays a significant role in the ischemia stage and reperfusion stage. Previous studies have indicated that both calcium/calmodulin-dependent protein kinase II (CaMKII) and glucose 6-phosphate dehydrogenase (G6PD) are involved in the oxidative stress. Thus, the aim of this study was to investigate the roles of CaMKIIα, an important isoform of CaMKII, and G6PD in a rat model of middle cerebral artery occlusion (MCAO). Intracerebroventricular injection of small interfering ribonucleic acid (siRNA) for CaMKIIα was performed at 48 h pre-MCAO surgery. Immunofluorescence Staining and western blot were performed to detect the expression of p-CaMKIIα and G6PD in the cortices. 2, 3, 5-Triphenyltetrazolium chloride (TTC) staining was performed to investigate the infarct volume. In addition, neurological deficit, reactive oxygen species (ROS), ratio of reduced-to-oxidized glutathione (GSH/GSSG) and ratio of reduced-to-oxidized oxidized nicotinamide adenine dinucleotide phosphate (NADPH/NADP+) were assessed. The results indicated that both p-CaMKIIα and G6PD were widely located in the neurons and astrocytes, and their expression was gradually increased in the cortices after MCAO, which was accompanied by increased level of ROS and decreased levels of GSH/GSSG and NADPH/NADP+. However, after treatment with siRNA for CaMKIIα, p-CaMKIIα expression was decreased and G6PD expression was increased. Moreover, inhibition of CaMKIIα improved the neurological deficit, reduced the infarct volume, decreased the level of ROS and increased the levels of GSH/GSSG and NADPH/NADP+. The results suggested that CaMKIIα inhibition exerted neuroprotective effects through regulating G6PD expression, which provides a new target for prevention and treatment of stroke.

Levels of Plasma N-terminal Pro-brain Natriuretic Peptide and D-dimer on the Prognosis of Patients with Acute Cerebral Infarction.

OBJECTIVE: To investigate the effects of levels of D-dimer and N-terminal pro-brain natriuretic peptide (NT-pro BNP) on the prognosis of patients with acute cerebral infarction. METHODS: One hundred and twenty-four patients with acute cerebral infarction who were admitted to the hospital between July 2014 and July 2016 were selected as the observation group; 100 normal people who had health examination in the center of physical examination of our hospital were selected as the control group. The levels of D-dimer and NT-pro BNP of the two groups were observed; the correlation between the levels of plasma NT-pro BNP and D-dimer and area of cerebral infarction, complications and death condition of the observation group was investigated. RESULTS: The levels of D-dimer and NT-pro BNP of the observation group were much higher than those of the control group, and the difference was statistically significant (P<0.05). The levels of D-dimer and NT-pro BNP of the observation group were significantly higher than those of the control group, and the difference had statistical significance (P<0.05). The levels of plasma NT-pro BNP and D-dimer of the patients with disturbance of consciousness and high blood pressure were apparently higher than those with no disturbance of consciousness and normal blood pressure, and there was a statistically significant difference (P<0.05). The patients were followed up for half a year. The levels of D-dimer and NT-pro BNP of the dead patients were much higher than those of the survived patients on admission. CONCLUSION: The levels of plasma NT-pro BNP and D-dimer can reflect the disease condition and prognosis of patients with acute cerebral infarction. Higher levels of NT-pro BNP and D-dimer indicates poorer prognosis. This work can provide a guidance for the clinical treatment of acute cerebral infarction.

Pratensein Mitigates Oxidative Stress and NLRP3 Inflammasome Activation in OGD/R-Injured HT22 Cells by Activating Nrf2-Anti-oxidant Signaling.

The current investigation seeks to uncover the neuroprotective effects and mechanisms of pratensein (Pra) against cerebral ischemia-reperfusion (CI/R) injury. An in vitro model was created by subjecting HT22 cells to oxygen-glucose deprivation/reoxygenation (OGD/R) injury. Various doses of Pra were administered to HT22 cells during the process of OGD/R. Nrf2 knockdown was achieved by siRNA transfection. Pra antagonized OGD/R-triggered HT22 cell damage, as suggested by increased cell viability and reduced levels of LDH secretion. Additionally, Pra reversed OGD/R-induced cell apoptosis, oxidative stress, and inflammatory injury. Transfection of Nrf2 siRNA partially ameliorated the protective effects of Pra on the OGD/R-stimulated increase in cell apoptosis, oxidative stress, and inflammatory response in HT22 cells. Pra significantly inhibited the expression of nod-like-receptor-protein-3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), caspase-1, and cleaved caspase-1 protein in OGD/R-induced cells. Nrf2 knockdown reversed the benefits of Pra on NLRP3 inflammasome activation. Besides, Pra administration mitigated middle cerebral artery occlusion/reperfusion-induced cerebral infarction, neurological deficits, and neuronal apoptosis in vivo. This study found that Pra suppresses NLRP3 inflammasome activation through Nrf2 activation, resulting in reduced inflammatory responses and rates of apoptosis in OGD/R-stimulated HT22 cells, highlighting the neuroprotective properties of Pra in CI/R.

Urban-rural difference in the costs of disability and its effects on poverty among people with disabilities in China.

The urban-rural difference in poverty is an important issue in China, particularly for people with disabilities. The extra costs of disability render this population susceptible to falling into poverty, where this can exacerbate the inequality among people with disabilities between urban and rural areas of the country. Previous studies have provided empirical evidence for the extra costs of disabilities in certain countries, but little scholarly attention has been devoted to the urban-rural gap in the costs of disability, particularly in countries like China that have a dual urban-rural system. This study explores changes in the extra costs of disability in China between urban and rural households with disabled members from 2008 to 2018 by using the standard of living approach. We apply the Foster-Greer-Thorbecke Poverty Index to measure the rates of poverty in urban and rural households with disabilities after considering the costs of disability. The results reveal that the costs of disability were not always lower for rural households than for urban households. At the same time, many rural households with disabled people were found to suffer from severe poverty owing to the high costs of their disabilities. The difference in health insurance and rehabilitation services between urban and rural China have led to an urban-rural gap in the costs of disability. This suggests that supplying more goods and services for disabled people in rural areas, especially free services, and raising the reimbursement due to them from their health insurance can help improve their standard of living.

[Correlation of the CTD structural domain of hepatitis B virus core protein with the encapsulation effect of indocyanine green].

Hepatitis B virus core protein (HBc) has become a hot spot in drug carrier protein research due to its natural particle self-assembly ability and ease of modification. The truncation of the C-terminal polyarginine domain (CTD, aa 151-183) of HBc does not affect the self-assembly of the particles. However, it does affect the internal and external charges of the particles, which may subsequently affect drug encapsulation. Thus, the truncated C-terminal polyarginine domain (CTD) of HBc and the inserted RGD peptide were selected to construct and express three HBc variants (RH) encapsulated with ICG (RH/ICG) with different C-terminal lengths to compare the stability and drug activity of their nanoformulations. RH160/ICG was found to have a great advantages in encapsulation efficiency and biological imaging. Compared with other HBc variants, RH160/ICG significantly improved encapsulation efficiency, up to 32.77%±1.23%. Cytotoxicity and hemolysis assays further demonstrated the good biocompatibility of RH160/ICG. Cell uptake and in vivo imaging experiments in mice showed that RH160/ICG could efficiently deliver ICG in tumor cells and tumor sites with good imaging effect. This research provides a new direction for further expanding the diagnosis and treatment application of ICG and development of HBc-based nanoparticle drug carrier platform.

Isolation and characterization of the first putative peroxidase gene from oilseed rape (Brassica napus) which is highly homologous to HRPC.

A new gene, designated as BnPrx (GenBank Accession No. DQ078754), was isolated from oilseed rape (Brassica napus) by SMART Rapid Amplification of cDNA Ends (RACE). The full-length cDNA is 1307 bp long and contains a 1062 bp open reading frame (ORF), which encodes a 354 amino acid peroxidase precursor, with a 31 aa N-terminal signal peptide and a 15 aa C-terminal propeptide. The putative protein has a molecular weight of 38.86 kDa and a calculated pI of 5.85. BnPrx shares high identity with HRPC (89%). BnPrx possesses all active residues and two Ca(2+) sites present in Horseradish peroxidase isoenzymes C (HRPC) as well as six N-glycosylation sites. The predicted 3-D structure of BnPrx is very similar to that of HRPC. Assisted by genomic walking technology, the genomic DNA of BnPrx was also cloned, consisting of 3 introns and 4 exons. Thirty-two TATA boxes, 18 CAAT boxes and many cis-elements, such as WUN, MeJR, were found in its promoter region. Southern blot analysis indicated that BnPrx belonged to a small gene family. Northern blot analysis revealed that BnPrx was constitutively expressed in all tested tissues, including roots, stems and leaves, with the high expression in leaves and stems. The expression of BnPrx could be induced by methyl jasmonate (MeJA), salicylic acid (SA), cold and H(2)O(2). The cloning and characterizing of BnPrx might not only help us understand the physiological function and molecular evolution of the large peroxidase gene family more comprehensively, but also provide an alternative way of seeking a more effective and economical substitute for HRPC.

[An intron-free methyl jasmonate inducible geranylgeranyl diphosphate synthase gene from Taxus media and its functional identification in yeast].

Geranylgeranyl diphosphate synthase (GGPPS, EC: 2.5.1.29) catalyzes the biosynthesis of geranylgeranyl diphosphate (GGPP), which is a key precursor for diterpenes including Taxol, one of the most potent antitumor drugs. In order to investigate the role of GGPP synthase in taxol biosynthesis, we cloned, characterized and functionally expressed the GGPP synthase gene from Taxus media. A 3743-bp genomic sequence of T. media was isolated by genome walking strategy which contained an 1182-bp open reading frame (ORF) encoding a 393-amino acid polypeptide that showed high similarity to other plant GGPPSs. Subsequently the full-length cDNA of the GGPPS gene of T. media (designated TmGGPPS) was amplified by RACE. Bioinformatic analysis showed that TmGGPPS was an intron-free gene and its deduced polypeptide contained all the five conserved domains and functional aspartate-rich motifs of the prenyltransferases. By constructing the phylogenetic tree of plant GGPPSs, it was found that plant-derived GGPPSs could be divided into two classes, angiosperm and gymnosperm classes, which might have evolved in parallel from the same ancestor. To our knowledge this was the first report that the geranylgeranyl diphosphate synthase genes were free of intron and evolved in parallel between angiosperms and gymnosperms. The coding sequence of TmGGPPS was expressed in yeast mutant (SFNY368) lacking of GGPP synthase activity through functional complementation, and the transgenic yeast showed to have activity of GGPP synthase. This was also the first time to use SFNY368 to identify the function of plant-derived GGPPSs. Furthermore, investigation of the impact of methyl jasmonate (MeJA) on the expression of TmGGPPS revealed that MeJA-treated T. media cultured cells had much higher expression of TmGGPPS than untreated cells.