AIM:To investigate the effect of AZD5363, an inhibitor of Akt, on the viability, apoptosis and autophagy in human hepatocelluar carcinoma cells and the molecular mechanisms.METHODS:MTT assay was used to detect the cell viability.TUNEL assay was used to analysis the apoptosis.The expression of PARP and LC3-II proteins was examined by Western blot analysis.The autophagy was characterized by the expression and distribution of GFP-LC3.RE-SULTS:The results of MTT assay indicated that AZD5363 suppressed the cell viability in a dose-dependent manner (P<0.05).High doses of AZD5363 triggered apoptosis via activating the cleavage of PARP.AZD5363 treatment induced autophagy both in Hep G2 cells and Huh7 cells by increasing the level of LC3-II.Blockage of autophagy by chloroquine promoted AZD5363-induced apoptosis in the hepatocellular carcinoma cells.CONCLUSION:AZD5363 increased apoptosis and autophagy in Hep G2 cells and Huh7 cells.Blockage of autophagy magnified AZD5363-induced apoptosis in hepatocellular carcinoma cells.

OBJECTIVETo investigate the effect of ketogenic diet (KD) on the clinical and electroencephalogram features in children with pharmacoresistant epileptic encephalopathy.

METHODThirty-one children (19 boys, 12 girls) aged 7 months to 7 years (mean 2 years 5 month) with epilepsy refractory to conventional antiepileptic drugs (AEDs) were included in this study. In addition to their original AED treatment, the children were assigned to different ketogenic diets based on their age. The prospective electro-clinical assessment was performed prior to the KD and then one week, one month and again 3 months after the initiation of therapy, respectively.

RESULTThe reduction of seizure frequency in 52%, 68% and 71% of all patients exceeded 50% one week, one month and three months after KD treatment respectively. KD is particularly effective in myoclonic astatic epilepsy (MAE; Doose Syndrome) and West syndrome with 100% and 81.25% of the patients having a greater than 50% seizure reduction, respectively. After 3 months of KD treatment, more than 2/3 patients experienced a reduction in interictal epileptiform discharges (IEDs) and improvement in EEG background.

CONCLUSIONThe clinical and electroencephalographic improvement confirms that KD is beneficial in children with refractory epilepsy.

Anticonvulsants ; therapeutic use ; Brain ; diagnostic imaging ; physiopathology ; Child ; Child, Preschool ; Diet, Ketogenic ; methods ; Dietary Fats ; administration & dosage ; Electroencephalography ; Epilepsy ; diagnosis ; diet therapy ; drug therapy ; Female ; Humans ; Infant ; Intellectual Disability ; diet therapy ; drug therapy ; Lennox Gastaut Syndrome ; Male ; Radiography ; Retrospective Studies ; Spasms, Infantile ; diet therapy ; drug therapy ; Syndrome ; Time Factors ; Treatment Outcome

OBJECTIVETo investigate therapeutic strategy of fulminant acute pancreatitis (FAP) in acute response stage.

METHODSSixty-four patients were divided into Death group (27 patients) and Survival group (37 patients). The time course of shock and recovery of enteral function, parameters of fluid resuscitation, PaO(2)/FiO(2) and AaDO(2) at 24 hours prior to mechanical ventilation, rate of continuous venovenous hemofiltration (CVVH) and abdominal compartment syndrome (ACS), severity of the disease in the acute response stage were investigated. And the effect of surgical manner and time on the prognosis was also analyzed.

RESULTSCompared with Survival group, the time course of shock and recovery of enteral function in Death group were prolonged significantly (P < 0.05). Between the groups, there was no difference in the amount of crystal fluid infused from admission to 72 hours after, but the amount of colloid fluid infused and ratio of amount of colloid and crystal fluid in Survival group were higher (P < 0.05). The amount of fluid retention in third space from admission to 72 hours after in Death group was higher than that of Survival group significantly (P < 0.05). The fluid infusing rate in Survival group in the first day of admission was faster than Death group (P < 0.05). PaO(2)/FiO(2) and AaDO(2) in 24 hours prior to mechanical ventilation in Death group were negatively changed significantly. Within 72 hours after the onset of the disease, the rate of CVVH in Survival group was higher than Death group. Incidence rate of ACS and the APACHEII scores within 72 hours after admission in Death group were higher than in Survival group. The cure rate of the patients operated in the day 7 to day 14 after admission was higher than that of patients operated prior and post this period. Time for the first operation in operated patients was earlier than patients received minimally invasive drainage (MID) and its cure rate was lower than that of MID Group.

CONCLUSIONSIt is the key point to shorten the time course of ischemia, to control persistent systemic inflammatory response syndrome (SIRS) and to adopt reasonable surgical intervention in acute response stage for FAP.

Acute Disease ; Adult ; Combined Modality Therapy ; Female ; Humans ; Male ; Middle Aged ; Pancreatitis ; complications ; mortality ; therapy ; Resuscitation ; methods ; Retrospective Studies

Objective· To investigate the relationship between pathological myopia and classification of vitreoretinal interface features using enhanced vitreous imaging optical coherence tomography (EVI-OCT). Methods · High myopia patients were included from 2015 to 2016. All participants underwent standardized medical interviews and ophthalmic examination. Results · The included eyes were divided into two groups of pathological myopia and simple high myopia based on myopic macular degeneration observed on fundus photography . There were four types of vitreoretinal interface changes demonstrated on EVI-OCT scans in included eyes: Type1, posterior precortical vitreous pockets (PPVP), Type2, partial posterior vitreous detachment with vitreous adhesion (VA), Type 3, epiretinal membrane (ERM), and Type 4, no traction (NT). Pathological myopia was mostly detected in VA, ERM, and NT groups. Conclusion · EVI-OCT was able to demonstrate the early changes of vitreoretinal interface in high myopia eyes. Vitreous adhesions and traction detected by OCT may facilitate the occurrence of pathological myopia.

OBJECTIVETo observe the pathological change of partial liquid ventilation (PLV) through establishing the rabbit model of acute lung injury (ALI) induced by meconium aspiration.

METHODSAdult, healthy male or female New Zealand white rabbits were randomly allocated into six groups as follows: (1) control group, (2) conventional mechanical ventilation (CMV) group, (3) high-frequency oscillatory ventilation (HFOV) group, (4) CMV combined with PLV group, (5) HFOV combined with PLV group and (6) normal group. The animals were anesthetized with 1% pentobarbital and tracheotomy was performed and endotracheal tube was placed, 20% meconium fluid (3 ml/kg) was quickly injected into the lung through the endotracheal tube and arterial blood gas was analyzed 30 minutes later. ALI was indicated when P/F ratio (PaO2)/FiO(2)) was < or = 300 mm Hg (1 mm Hg = 0.133 kPa) and Cdyn Dynamic Compliance declined by more than 30% of the baseline. The animals were then randomly allocated into one of the 6 groups. In PLV groups (including CMV + PLV and HFOV + PLV) warmed (37 degrees C) and oxygenated perfluorocarbon was slowly instilled into the lungs of the rabbits through the endotracheal tube at a low-dose 3 ml/kg, then set 15-min positive pressure by sacculus proprius to guarantee perfluorocarbon to steadily diffuse in to the lungs. Six hours after ventilation the animals were sacrificed by using overdose of room air instillation via vein. The lungs were taken and fixed in 4% paraformaldehyde (PFA) and were stained with hematoxylin-eosin (HE). Pathological evaluations included inflammatory manifestation, edema and hemorrhage in both alveolar and interstitial area, damages of small airway (alveolar tube and alveolar bursa) and hyaline membrane formation. One way analysis of variance, Student Newman-Keuls (SNK) method and Kruskal-Wallis (K-W) test were used for comparisons.

RESULTSWith the exception of normal group 30 minutes after meconium injections blood gas analysis in different groups showed significant changes and PaO(2)/FiO(2) (< 300 mm Hg), Cdyn declined by more than 60% compared with baseline (P < 0.05). The pathological analysis showed that alveolar and interstitial inflammation, edema, alveolar and interstitial hemorrhage, and small airway damage existed in each group. The hyaline membrane formation was found in one of CMV + PLV group rabbits. The perfluorocarbon-treated animals (CMV + PLV and HFOV + PLV) showed significantly less injury in dependent lung and less damage of small airway (CMV + PLV or HFOV + PLV vs. CMV = 1.1 +/- 0.4 or 0.9 +/- 0.3 vs 2.6 +/- 0.5) compared with the animals of CMV group (P < 0.01). HFOV group (2.1 +/- 0.3) also had less alveolar and interstitial inflammation compared with CMV group (3.0 +/- 0) (P < 0.05), and there was less evidence of alveolar and interstitial edema in the animals treated with HFOV + PLV (1.0 +/- 0.7) compared with CMV (2.0 +/- 0.8) (P < 0.01). Treatment with perfluorocarbon did not result in significant difference in alveolar and interstitial hemorrhage. Compared with CMV and HFOV groups, the groups treated with PLV showed lower mortality of animals (21.4% and 14.3%).

CONCLUSIONSPLV can alleviate the histological damage of acute lung injury induced by meconium aspiration and increased survival chance and therefore PLV would be a useful treatment for MAS. The effectiveness and safety of application of PLV should be evaluated in clinical studies.

Acute Lung Injury ; etiology ; pathology ; Animals ; Animals, Newborn ; Disease Models, Animal ; Female ; Liquid Ventilation ; Male ; Rabbits

Coronary Angiography ; Coronary Vessel Anomalies ; diagnostic imaging ; therapy ; Embolization, Therapeutic ; methods ; Fistula ; congenital ; diagnostic imaging ; therapy ; Heart Ventricles ; abnormalities ; diagnostic imaging ; Humans ; Male ; Middle Aged

Objective To study the effect of human umbilical cord mesenchymal stem cells (hUCMSCs) tail vein transplantation on depressive behavior in chronic unpredictable mild stress (CUMS) model mice. Methods The third generation hUCMSCs were abtained; Sixty C57BL/6 male mice were randomly divided into the normal model group (+ normal saline), the cell group (+hUCMSCs) and the fluoxetine(flu) group (+flu), with 15 mice in each group. The depression model of CUMS mice with a 6 week duration was constructed; From the third week, hUCMSCs and normal saline were transplanted into the mice by tail vein injection, and the mice were gavaged with flu daily from the fifth week; Weight changes in each group were recorded weekly; The depression model and therapeutic effect of mice were evaluated by sucrose preference test, tail suspension test, forced swimming test and open field test; The cell changes of CAI and CA3 region in hippocampus were observed by HE staining; Transmission electron microscopy was used to detect the changes of synapses in CA3 region of hippocampus; Real-time PCR was used to detect mRNA expressions of synaphysin (SYP) and postsynaptic density protein 95 (PSD-95), which are key proteins of synaptic plasticity. Results hUCMSCs improved weight decrease in depressed mice; behavioral experiments observed that the model group mice showed depressive behavior, while the cell group and the drug group mice depression were improved; HE staining showed that, compared with the model group, cells in CA 1 and CA3 regions of the hippocampus of mice in the cell group and the group were arranged orderly and the number of cells increased; Transmission electron microscopy showed that compared with the model group, the number of synapses in the CA3 region of the hippocampus was more, the synaptic gap was narrower, and the density of synaptic spines was higher in the cell group and the drug group; More mRNA expressions of SYP and PSD-95 were detected by Real-time PCR in the cell group and the drug group than in the model group. Conclusion hUCMSCs transplatation showes antidepressant effects, which are associated with improved synaptic plasticity in the hippocampus.

Animals ; Cellular Reprogramming ; Dentate Gyrus ; cytology ; Fibroblasts ; cytology ; Mice ; Neural Stem Cells ; cytology ; transplantation ; Swine

Objective:To observe the effect of isopimpinellin on primary hippocampal neuron cells γ-aminobutyric acid (GABA), 5-hydroxytryptamine (5-HT) and receptor genes expressions, in order to explore its hypnotic mechanism. Method:The primary hippocampal neurons of neonatal Sprague-Dawley rats were cultured in vitro. And subsequent experiments were conducted in the optimal state of cell growth, and the purity was identified by immunohistochemistry of neuron-specific enolase. Hippocampal neurons were randomly divided into five groups, namely blank control group, diazepam group (25 mg·L^-1), and low-dose (5 mg·L^-1), moderate-dose (10 mg·L^-1) and high-dose (20 mg·L^-1) isopimpinellin groups. Early apoptosis of hippocampus neuron cells were detected using flow cytometry technique after 24 h administration, and the changes in the levels of GABA and 5-HT were detected using enzyme-linked immunosorbent. The changes in mRNA expressions of receptor genes relating to gamma-aminobutyric acid type A receptor(GABAA) genes GABRA₁,GABRA₅,GABBR₁, gamma-aminobutyric acid type B receptor genes (GABAB) GABRB₂, 5-hydroxytryptamine 1A receptor (5-HT1_A)5-HT1_A(A),5-HT1_A(B),5-HT1_A(C) were detected by real-time quantitative PCR(Real-time PCR). Result:On the 7^th day, the hippocampal neurons grew in a good condition, and the purity was above 90%. Apoptosis rates of hippocampal neurons in the low-dose and moderate-dose groups were significantly lower than that in the blank control group (P<0.01). The level of GABA secreted by hippocampal neurons in the high-dose isopimpinellin group were significantly higher than that in the blank control group (P<0.01). The mRNA expression levels of GABRA₁,GABRA₅,5-HT1_A(A),5-HT1_A(C) in the moderate-dose and high-dose isopimpinellin groups were significantly higher than those in the blank control group (P<0.05, P<0.01). The mRNA expression levels of GABBR₁,5-HT1_A(B) in the low-dose, moderate-dose and high-dose isopimpinellin groups were significantly higher than those in the blank control group (P<0.05, P<0.01). Conclusion:The hypnotic mechanism of isopimpinellin may be related to the inhibition of hippocampal neuron apoptosis, the increase of the content of inhibitory neurotransmitter GABA, and the up-regulation of GABA and 5-HT-related receptor genes.

Objective:To observe the expression of brown adipose tissue (BAT), cells, proteins and corresponding genes in Yang deficiency model mice induced by Rhei Radix et Rhizoma suspension, and to explore the thermogenesis of processed products of Aconiti Lateralis Radix Praeparata with Jianchang faction characteristics. Method:Twenty mice, half male and half female, were randomly selected as the normal female and male groups. And the other 80 mice were administrated with Rhei Radix et Rhizoma suspension (the content of 0.25 g·mL^-1) to establish Yang deficiency model, after the model was established, they were randomly divided into the model female and male groups, female and male groups of Shengfupian, female and male groups of Yinfupian, female and male groups of Yangfupian, 10 mice in each group. Mice were intragastric administrated with corresponding medical solution for two weeks (1.54 g·kg^-1·d^-1) according to groups. Normal group and model group were given equal volume distilled water. After administration, BAT of scapular region of mice was collected and the changes of BAT cells were observed by hematoxylin-eosin (HE) staining. The expression of uncoupling protein 1 (UCP1) and its mRNA were detected by Western blot and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Result:Compared with the normal group of the same sex, the proportion of BAT in the model group decreased significantly (P<0.01). Compared with the model group of the same sex, the proportion of BAT in female mice from Shengfupian and Yinfupian groups increased significantly (P<0.01), while there was no significant difference between each administration group and model group in the male mice. Compared with normal mice of the same sex, there were many scattered vacuoles in BAT cells of the model group, and fewer cells could be observed due to larger vacuoles. Compared with the model group of the same sex, BAT cells in mice from the Shengfupian group showed fewer vacuoles, smaller cells and tight arrangement, the density of BAT cells in mice from the Yangfupian group also increased significantly, while the vacuoles in BAT cells of mice from the Yinfupian group decreased relatively and the cells did not increase significantly. Compared with the same sex mice, the expression level of UCP1 in the model group and the normal group was statistically significant (P<0.05, P<0.01). In the female mice, the expression level of UCP1 in Yangfupian group was significantly higher than that in the model group (P<0.05), each administration group of male mice was significantly different from that of the model group of the same sex (P<0.05), of which Yangfupian was the most significant. The relative expression of UCP1 mRNA in the model group was significantly lower than that in the normal group of the same sex (P<0.05, P<0.01). In the female mice, compared with the model group, the relative expression levels of UCP1 mRNA in Yangfupian group, Shengfupian group and Yinfupian group increased significantly (P<0.05, P<0.01), compared with Yangfupian group, the relative expression levels of UCP1 mRNA in Shengfupian and Yinfupian were also significantly different (P<0.05). In the male mice, compared with the model group, the relative expression of UCP1 mRNA in Yangfupian group was significantly increased (P<0.01), but there was no significant difference in Shengfupian group and Yinfupian group, in addition, compared with Yangfupian group, the relative expression of UCP1 mRNA in Shengfupian group and Yinfupian group had significant difference (P<0.05). Conclusion:Shengfupian, Yinfupian and Yangfupian all have obvious improvement on Yang deficiency syndrome induced by Rhei Radix et Rhizoma suspension. The mechanism may be to promote the expression of UCP1 protein and its mRNA and enhance the activity of BAT. And the effect of Yangfupian is the best.