Distribution of phosphorylated Elk-1 in rat brain after Y-maze active avoidance training in a temporal manner

BACKGROUND： Elk-1 mRNA distributes extensively in the neurons of mice, rat and human brains, and the Elk-1 expression may be correlated with the synaptic plasticity, learning and memory. OBJECTIVE： To observe the distribution of phosphorylated Elk-1 （pEIk-1） in whole brain of rats received Y-maze active avoidance training and the changes of pEIk-1 expression at different time points after training. DESIGN ： A randomized controlled study SETTING ： Research Room of Neurobiology, the Second Affiliated Hospital of Southern Medical University MATERIALS ： Fifty-five male clean-degree SD rats of 3-4 months old, weighing 200-250 g, were provided by the Experimental Animal Center of Southem Medical University. The rabbit anti-monoclonal pEIk-1 antibody was purchased from Cell Signal Transduction Company, and ABC kit from Vector Company. METHODS ： The experiments were carried out in the Research Room of Neurobiology, Second Affiliated Hospital of Southern Medical University from September 2004 to February 2005. ① Grouping： The rats were randomly divided into training group （n = 25）, sham-training group （n = 25） and normal control group （n = 5）, and the training and sham-training groups were observed at 0, 1, 3, 6 and 24 hours after training, which represented the five phases in the process of leaming and memory. ② Y-maze training： The rats were preconditioned in the electrical Y-maze apparatus, 20 minutes a day for 3 days continuously, and training began from the 4^th day. In the training group, the rats were trained with the combination of light and electddty. Each rat repeated for 60 times in each training, and the correct times were recorded, those correct for less than 25 times were taken as unqualified, and excluded from the training group, and supplemented by other rats in time. In the sham-training group, there was no fixed correlation between the application of light and electricity. The rats in the normal contrel group were given not any training. ③Detection of pEIk-1 expression： The rats were anesthetized after Y-maze training, brain tissue was removed to prepare coronal freezing sections, and the pEIk-1 expression was detected with routine ABC method. MATN OUTCOME MEASURES： ① Distribution of pEIk-1 immuno-positive neurons in whole brain of rats in the normal control group. ②Comparison of the expression of pEIk-1 immuno-positive neurons in whole brain at different time points after training between the training group and sham-training group. RESULTS ： All the 55 rats were involved in result analysis. ③ Distribution of pEIk-1 immuno-positive neurons in the whole brain of rats in the normal control group： Strong expressions of pEIk-1 immuno-positive neurons were observed in prefrontal lobe, granular layer of olfactory bulbs, Purkinje cell layer and granular layer of cerebellum, whole stdate cortex, temporal cortex, pre-pyriform cortex, hypothalamic supraoptic nucleus, hypothalamic paraventricular nucleus and periventricular nucleus, thalamic paraventricular nucleus, pronucleus and postnucleus of amygdala cortex, central nucleus of amygdala, medial amygdaloid nucleus, entorhinal cortex, hippocampal dentate gyros, CA1-4 regions, caudate-putamen, material division, brain stem spinal nucleus of trigeminal nerve, and superior olivary nucleus, and those in hippocampal dentate gyrus and CA1 region were the strongest.② Distribution of pEIk-1 immuno-positive neurons in the whole brain of rats at different time points after training in the training group and sham-training group： In the training group, the expressions were obviously enhanced in caudate-putamen of striatum, material division, most cortexes, hippocampal dentate gyrus, hippocampal CA regions, nucleus amygdalae, thalamic paraventricular nucleus, Purkinje cell layer of cerebellum, entorhinal cortex, hypothalamic supraoptic nucleus, hypothalamic paraventricular nucleus, and periventricular nucleus at 0 hour after training, and the enhancement lasted for 6 hours at least, and those at 24 hours were decreased to normal. In the sham-training group, obvious enhanced expressions of pEIk-1 immuno-positive neurons could be observed in most cortexes, nucleus amygdalae, entorhinal cortex, hypothalamic supraoptic nucleus, hypothalamic paraventdoular nucleus and periventricular nucleus, brain stem spinal nucleus of trigeminal nerve, Purkinje cell layer and granular layer of cerebellum at O, 1, 3 and 6 hours, and decreased to normal after 24 hours. The expressions in material division, caudate-putamen of striatum, hippocampus were not obviously enhanced as compared with those in the normal control group, but significantly different from those in the training group （0, 1, 3, 6 hours after training, material division： F= 0.576, 0.023, 0.116, 8.873, P〈 0.01; caudate-putamen： F= 0.157, 0.427, 0.030, 0.001, P〈 0.01; hippocampus： F= 6.716, 2.405, 14.137, 1.416, P 〈 0.05-0.01 ）. CONCLUSION： The expression of activated pEIk-1 can be detected in the learning related brain areas under normal status, and the perk-1 expression in the brain areas dynamically changed in a time-dependent manner after Y-maze training, and it is indicated that pEIk-1 is involved in the learning and memory process in Y-maze related brain areas.

Rapid construction of phosphatase and tensin homolog-deleted on chromosome ten gene recombinant adenovirus using the AdEasy system

Recent studies have shown that phosphatase and tensin homolog-deleted on chromosome ten （PTEN） gene plays an important role in ischemic brain damage and synaptic plasticity. The AdEasy system, which has been widely used, greatly simplifies preparation of recombinant adenovirus. Therefore, recombinant defective adenovirus vector carrying human PTEN tumor suppressor gene （Ad-PTEN） was constructed using the AdEasy-1 system and was transfected into HEK293 cells for packaging and amplification. Infection efficiency and expression intensity were observed in primary cultured rat hippocampal neurons infected with Ad-PTEN in vitro. Results revealed a cytopathic effect in green fluorescent protein expression, which increased with prolonged time. After three cycles of amplification, the adenovirus titer was increased to an adequate titer for infecting hippocampal neurons. The entire process typically requires 4-5 weeks for completion. Results suggested that recombinant defective adenovirus vector carrying the PTEN gene was successfully and rapidly constructed using the AdEasy system.

Impacts of Opinion Propagation on Social Balance

In social networks, opinions diffusion often leads to relationships evolution. Then changes of relationships result in the change of balance degree of social system. We simulate the opinion diffusion on Barabasi & Albert (BA) network and Watts & Strogatz (WS) network to study the effects of the two types of networks, dynamical parameters and structural parameters on the balance degree of system. We employ the spectral analysis to quantify the balance degree of system before and after opinion propagation. The result reveals that it is very similar effect of BA networks and WS networks on it. However, it is opposite effects between dynamical parameters and structural parameters. The balance degree of system is proportional to the two dynamical factors (P,Q) at initial state and always inversely proportional to the two structural factors (,Pne) at initial and convergence state.

Impact of vaccination against COVID-19 on the outcomes of in vitro fertilization–embryo transfer:a retrospective cohort study

Background: Vaccination against coronavirus disease 2019 (COVID-19) has become the primary approach in the fight against the spread of COVID-19. Studies have shown that vaccination against COVID-19 has adverse effects, particularly on human reproductive health, despite the fact that vaccination rates are still on the rise. However, few studies have reported whether vaccination affects the outcome of in vitro fertilization-embryo transfer (IVF-ET) or not. In this study, we compared the outcome of IVF-ET and the development of follicles and embryos between vaccinated and unvaccinated groups. Methods: A single-center retrospective cohort study of 10,541 in vitro fertilization (IVF) cycles was conducted from June 2020 to August 2021. 835 IVF cycles with a history of vaccination against COVID-19 and 1670 IVF cycles that served as negative controls were selected and analyzed utilizing the Matchlt package of R software (http://www.R-project.org/) and the nearest neighbor matching algorithm for propensity-matched analysis at a 1:2 ratio. Results: The number of oocytes collected in the vaccinated group and the unvaccinated group were 8.00 (0, 40.00) and 9.00 (0, 77.00) ( P = 0.073) and the good-quality embryo rates of the two groups were 0.56±0.32 and 0.56±0.31 averagely ( P = 0.964). Clinical pregnancy rates for the vaccinated group and unvaccinated group were 42.4% (155/366) and 40.2% (328/816) ( P = 0.486) and biochemical pregnancy rates were 7.1% (26/366) and 8.7% (71/816) ( P = 0.355). Two other factors were analyzed in this study;vaccination among different genders and different types (inactivated vaccine or recombinant adenovirus vaccine) showed no statistically significant effect on the above outcomes. Conclusions: In our findings, vaccination against COVID-19 showed no statistically significant effect on the outcomes of IVF-ET and the development of follicles and embryos, nor did the gender of the vaccinated person or the formulation of vaccines show significant effects.

Improvement of antibacterial, anti-inflammatory, and osteogenic properties of OGP loaded Co-MOF coating on titanium implants for advanced osseointegration

The bacterial infection,especially for methicillin-resistant Staphylococcus aureus(MRSA),and the associated severe inflammatory response could extremely limit the crosstalk of RAW264.7 cells and mesenchymal stem cells(MSCs)and lead to the undesirable osseointegration of peri–implants.It is highly demanded to modify the surface of titanium(Ti)implant to improve its anti-bacterial and anti-inflammatory properties and facilitate its disabled osseointegration.Herein,in our study,a multifunctional coating of zeolitic imidazolate frameworks-67 encapsulated osteogenic growth peptide(OGP)(ZO)was fabricated on titanium dioxide nanotubes(TNT)substrates(TNT-ZO)via the electrophoresis deposition(EPD)approach.The TNT-ZO substrates exhibited excellent antibacterial activity indicated by the reactive oxygen species(ROS)generation,outer membrane(OM)and inner membrane(IM)permeabilization change,adenosine triphosphate(ATP)decrease,and intracellular compounds(DNA/RNA)leakage.Importantly,the regulation effects of TNT-ZO coating modified titanium substrates on the RAW264.7-MSCs crosstalk could induce the anti-inflammatory and osteogenic microenvironment via multiple paracrine signaling of Runx2,BMP2,VEGF,and TGF-β1.The promoted effects of coating structure were investigated in vivo,including antibacterial effect,osteogenic differentiation of mesenchymal stem cells,and anti-inflammation of RAW264.7 cells,as well as infected bone regeneration and repair in bone defect transplantation model.The results demonstrated that MRSA was effectively eliminated by the hydrolysis of ZIF-67 nanoparticles on TNT-ZO substrates.Furthermore,the excellent osseointegration of peri–implants was realized simultaneously by modulating the crosstalk of RAW264.7-MSCs.This study could provide a novel approach to designing a multifunctional coating on the Ti implants for infected bone regeneration in orthopedic applications.