Objective To observe the morphology of neuronal necrosis by kainic acid(KA) induced status epilepsy (SE) in rats, and to study the brain protective effect of Mg 2+.Methods 75 adult male Wistar rats were randomly divided into KA group, Mg 2+ group and normal saline group. SE was induced with KA for 3 hours, and the rats in Mg 2+ group were intraperitoneal injected magnesium sulfate before being injected KA. 72 hours later the rats were killed. We had all rat brain sections and observe the morphology of neuronal necrosis with microscope and electron microscope.Results In KA group, seizure was induced 16.1?4.7 min after injection of KA, but seizure delayed to 25.4?6.2 min in Mg 2+ group. There was a significant difference between two groups (P

AIM:To investigate the time course of nuclear factor-κB (NF-κB) and the effects of 3-aminobenzamide (3-AB) on the expressions of NF-κB,interleukin-1β (IL-1β) and cyclooxygenase-2 (COX-2) in hippocampus after seizures. METHODS:Epilepsy were induced by kainic acid through cerebral ventricular injection. Western blotting was used to detect NF-κB p65 expression in nucleus at various experiment groups. Moreover,mRNA and protein expressions of IL-1β and COX-2 in different experiment groups were determined by RT-PCR and Western blotting analysis. RESULTS:NF-κB p65 immunoreactivity began to increase in the nuclear fraction at 2 h (P<0.05),kept rising at 12 h (P<0.05) and returned to control level at 24 h after epilepsy seizures. Furthermore,3-AB sharply decreased the accumulation of NF-κB p65 in nucleus (P<0.05). In addition,3-AB significantly decreased the mRNA and protein expressions of IL-1β and COX-2 which obviously increased in hippocampus at 6 h after epilepsy seizures (P<0.05). CONCLUSION:Seizures triggers NF-κB nucleus translocation and promotes the expressions of IL-1β and COX-2 in hippocampus. In addition,poly (adenosine diphosphate-ribose) polymerase inhibition by 3-AB suppresses NF-κB associated inflammatory pathway in epileptic rat hippocampus.

Objective To establish a primary culture method of hippocampal neurons of new-born rats,and to observe the morphological characteristics at different developmental and differential stages.Methods The hippocampus was digested and the cells were seeded in a flask.The neurons were then transferred and re-seeded on cover glasses coated with poly-L-lysine.The neurons were identified by polyclonal antibody against neuron specific enolase(NSE).The morphology was observed under phase-contrast microscope.Results A large number of hippocampal neurons began to adhere to the cover glasses after 12～24 hours.They showed different shapes-shuttle,triangle,pyramidal,or nonregular after clinging to the plate.Their processes connected into nets and different in length and thickness.They were well developed on the 7th～10th day and survived as long as 28 days after seeding.Immunocytochemistry of NSE proved high purity.Conclusion The culture method of new-born rat hippocampal neurons in vitro is successful and can be used as an in vitro model of research.

Objective To study the role of gap junctions in epileptiform activity. Methods The epileptiform activity was induced by zero-Mg 2+ medium in cultured hippocampal neurons of newborn rats. Immunocytochemistry and real time RT-PCR were introduced to evaluate the expression of gap junction Cx32 and Cx43. Results The level of Cx32 mRNA increased quickly one hour after the neurons were treated with zero-Mg 2+ medium and was raised by 10 times 5 hours later, while Cx32 protein began to develop at the 2nd hour (21.80?1.74) and was raised by 5 times at the 8th hour (47.30?5.75). The expression of Cx43 mRNA went up obviously 5 hours later, and Cx43 protein developed visibly 8 hours later. Carbenoxolone depressed the expressions of Cx32 and Cx43. Conclusions The expression of Cx32 and Cx43 increases dramatically after epileptic discharges and carbenoxolone inhibits both the discharges and the expression of gap junctions, which indicates that gap junction could contribute to epileptogenesis.

AIM:To investigate the time course of nuclear factor-?B (NF-?B) and the effects of 3-aminobenzamide (3-AB) on the expressions of NF-?B,interleukin-1? (IL-1?) and cyclooxygenase-2 (COX-2) in hippocampus after seizures. METHODS:Epilepsy were induced by kainic acid through cerebral ventricular injection. Western blotting was used to detect NF-?B p65 expression in nucleus at various experiment groups. Moreover,mRNA and protein expressions of IL-1? and COX-2 in different experiment groups were determined by RT-PCR and Western blotting analysis. RESULTS:NF-?B p65 immunoreactivity began to increase in the nuclear fraction at 2 h (P

Objective:To explore the CT three-dimensional reconstruction in percutaneous balloon compression of the semilunar ganglion of trigeminal nerves.Methods:Thirty-one patients with trigeminal neuralgia, admitted to our hospital from May 2018 to August 2018, were treated by percutaneous balloon compression of the semilunar ganglion of trigeminal nerves. All patients underwent intraoperative mobile CT scan; the distance from the external aperture of foramen ovale to the petrous ridge, the angle from puncture needle and the petrous ridge, and the angle from puncture needle and the clivus were analyzed by three-dimensional reconstruction technology. Prognoses were recorded immediately after surgery and 10-12 months after follow-up.Results:The distance from the external aperture of foramen ovale to the petrous ridgea was (2.14±0.17) cm, ranged from 1.83 cm to 2.42 cm. The angle from puncture needle and the petrous ridge was (102.03±7.60)°, ranged from 91.00° to 116.00°. The angle from puncture needle to the clivus was (68.03±7.06)°, ranged from 52.00° to 80.00°. The puncture succeeded in all patients under the help of CT three-dimensional reconstruction. The volume of the inflated balloon ranged from 0.30 cm 3 to 0.47 cm 3 (averaged 0.42±0.05 cm 3). The compression time ranged from 3.00 min to 5.00 min (averaged 4.72±0.53 min). The incidence of diplopia was 3.23%, and the incidence of dry eyes/decreased corneal sensation was 12.90%. During the follow-up, one patient relapsed 10 months after surgery; the left patients recovered with disappeared complications. Conclusion:The CT 3D reconstruction can reduce the difficulty of ovale foramen puncture during operation, assist the judgment and adjustment of intraoperative balloon volume, and improve the surgical efficacy.

Objective To prepare polymersomes (PSs) by block copolymers，evaluate their membrane structural stability，investigate the H⁺ transmembrane permeability of PSs and the impact of 1,4-dioxane and establish a foundation for drug encapsulation within polymersomes. Methods PSs were self-assembled by a block copolymer, PEG-PLGA, in a solvent solution. The pH-sensitive fluorescence probe HPTS was employed to examine the H⁺ transmembrane properties of PSs and compare them with PSs prepared using PBD-b-PEO, PS-b-PEO, and liposomes. The effect of varying concentrations of 1,4-dioxane on PSs’ membrane permeability properties was also investigated. Results The fluorescence excitation spectra of HPTS exhibited pH dependency, which showed a linear correlation between extravesicular H⁺ concentration and t^1/2. Significant differences were observed in the membrane permeability capabilities of PSs with different membrane wall thicknesses. Compared to liposomes, the H⁺ transmembrane coefficients for the three types of PSs were reduced by 2.39×10⁴, 3.38×10⁴, and 5.48×10⁸ times, respectively. 1,4-dioxane was found to modulate the permeability of PSs’ membranes, which displayed a concentration-dependent relationship. Conclusion PSs exhibited significantly lower membrane permeability compared to liposomes, indicating superior stability. 1,4-dioxane was identified as a modulator of PSs’ permeability, which offered potential for drug loading and release within PSs.

Objective To prepare and optimize the formulation of Albumin nanoparticles loading PROTAC molecule and observe the inhibition effect of nanoparticles on the proliferation and NAD⁺ metabolism of glioma cells. Methods Albumin nanoparticles loading NPT-B2 were prepared and characterized with a thermal driving method, and the prescription was optimized. An HPLC method was established to determine the content of NPT-B2. The proliferation inhibition of NPT-B2 and B2-BSA-NPs on U251 cells were investigated by the CCK8 method, and the degradation effects of NPT-B2 and B2-BSA-NPs on NAMPT in glioma cells were investigated by western blotting. Results The HPLC method was stable, with good linearity, precision, and recovery rate. The nanoparticles had a particle size of about 55.48 nm, a potential of about −12.9 mV, an encapsulation rate of about 94.74%, and a drug loading amount of about 8.61%. The IC₅₀ of NPT-B2 on glioma cells was 61.16 nmol/L, which had a degradation effect on NAMPT. The IC₅₀ of B2-BSA-NPs on glioma was 41.21 nmol/L, which had a very significant degradation effect on NAMPT. Conclusion Albumin nanoparticles loading PROTAC molecules were constructed. The prescription was optimized to improve the drug encapsulation rate, and the low water solubility of PROTAC molecule was improved, which had a significant inhibitory effect on the proliferation and NAD⁺ energy metabolism of glioma cells.