Physiological and lipidomic response of exogenous choline chloride alleviating salt stress injury in Kentucky bluegrass (Poa pratensis).

Introduction: Choline participates in plant stress tolerance through glycine betaine (GB) and phospholipid metabolism. As a salt-sensitive turfgrass species, Kentucky bluegrass (Poa pratensis) is the main turfgrass species in cool-season areas. Methods: To improve salinity tolerance and investigate the effects of choline on the physiological and lipidomic responses of turfgrass plants under salinity stress conditions, exogenous choline chloride was applied to Kentucky bluegrass exposed to salt stress. Results: From physiological indicators, exogenous choline chloride could alleviate salt stress injury in Kentucky bluegrass. Lipid analysis showed that exogenous choline chloride under salt-stress conditions remodeled the content of phospholipids, glycolipids, and lysophospholipids. Monogalactosyl diacylglycerol, digalactosyl diacylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and lysophosphatidylcholine content were increased and phosphatidic acid content were decreased in plants after exogenous choline chloride under salt treatment. Plant leaf choline content increased, but GB was not detected in exogenous choline chloride treatment plants under nonstress or salt-stress conditions. Discussion: GB synthesis pathway related genes showed no clear change to choline chloride treatment, whereas cytidyldiphosphate-choline (CDP-choline) pathway genes were upregulated by choline chloride treatment. These results reveal that lipid remodeling through choline metabolism plays an important role in the salt tolerance mechanism of Kentucky bluegrass. Furthermore, the lipids selected in this study could serve as biomarkers for further improvement of salt-sensitive grass species.

Identification and pharmacological characterization of histamine-gated chloride channels in the fall armyworm, Spodoptera frugiperda.

Histamine-gated chloride channels (HACls) mediate fast inhibitory neurotransmission in invertebrate nervous systems and have important roles in light reception, color processing, temperature preference and light-dark cycle. The fall armyworm, Spodoptera frugiperda is a main destructive pest of grain and row crops. However, the pharmacological characterization of HACls in S. frugiperda remain unknown. In this study, we identified two cDNAs encoding SfHACl1 and SfHACl2 in S. frugiperda. They had similar expression patterns and were most abundantly expressed in the head of larvae and at the egg stage. Electrophysiological analysis with the two-electrode voltage clamp method showed that histamine (HA) and γ-aminobutyric acid (GABA) activated inward currents when SfHACls were singly or collectively expressed with different ratios in Xenopus laevis oocytes. These channels were ≥2000-fold more sensitive to HA than to GABA. They were anion-selective channels, which were highly dependent on changes in external chloride concentrations, but insensitive to changes in external sodium concentrations. The insecticides abamectin (ABM) and emamectin benzoate (EB) also activated these channels with the EC50 to SfHACl1 lower than that to SfHACl2. And the EC50s of ABM and EB to the co-expressed channels gradually increased with increase in the injection ratio of SfHACl2 cRNA. Homology models and docking simulations revealed that HA bound to the large amino-terminal extracellular domain of SfHACl1 and SfHACl2 by forming 4 and 2 hydrogen bonds, respectively. The docking simulations of ABM and EB had similar binding sites in the transmembrane regions. Overall, these findings indicated that HACls act as targets for macrolide, and this study provides theoretical guidance for further derivatization of abamectin insecticides.

An ultra-sensitive and selective nitrogen dioxide sensor based on a novel P2C2 monolayer from a theoretical perspective.

The sensing properties of an α phase black phosphorus carbide (P2C2) monolayer for the adsorption of CO2, H2, H2O, N2, H2S, NH3, O2 and NO2 gases are theoretically investigated using first-principles calculations. We calculate the adsorption energy, equilibrium distance, Mulliken charge transfer, electron localization function, and work function to explore whether P2C2 is suitable for detecting NO2 gas. The results demonstrate that the P2C2 monolayer is highly sensitive and selective to NO2 gas molecules with robust adsorption energy and superior charge transfer due to the existence of strong orbital hybridization between the NO2 molecule and monolayer P2C2. In addition, the results of the work function calculations indicate that field effect transistor type NO2 gas sensors based on P2C2 monolayers are also feasible. Furthermore, the current-voltage curves reveal that the adsorption of NO2 can greatly modify the resistance of the P2C2 monolayer. Our results show that gas sensors based on P2C2 monolayers could be better than those based on black phosphorene (BP) for detecting NO2 molecules in an air mixture. In addition, the recovery time of the P2C2 sensor at T = 300 K was estimated to be short (and even shorter at higher temperatures) for NO2 which satisfies the demands for sustainable use.

Total arch repair with open triple-branched stent graft placement for acute type A aortic dissection: experience with 122 patients.

OBJECTIVE: To summarize the clinical experiences and midterm follow-up results of total arch repair with open triple-branched stent graft placement for acute type A aortic dissection. METHODS: From June 2008 to March 2013, 122 patients (95 men and 27 women; mean age, 50.9 ± 10.4 years) with acute type A aortic dissection underwent total arch repair with open placement of a triple-branched stent graft under hypothermic cardiopulmonary bypass and selective cerebral perfusion. During the follow-up period, enhanced computed tomography and echocardiography were performed at 3 months postoperatively and annually thereafter. RESULTS: Placement of the triple-branched stent graft into the true lumen of the descending aorta, arch, and 3 arch vessels was technically successful in 121 patients. The cardiopulmonary bypass time was 186.50 ± 38.23 minutes, and the selective antegrade cerebral perfusion time was 31.97 ± 10.08 minutes. The in-hospital mortality was 4.93%. No permanent neurologic dysfunction or paraplegia was observed. Three patients were lost to follow-up. The mean follow-up period was 30.24 ± 12.35 months. After hospital discharge, 3 patients died. On the 3-month postoperative scans, complete thrombus formation around the triple-branched stent graft was observed in 89.38% of the patients. Endoleaks were detected in 12 patients; 8 patients refused any management for the endoleaks, but they maintained a good quality of life. The other 4 patients were successfully treated by additional surgery. CONCLUSIONS: Total arch repair with open triple-branched stent graft placement is an effective technique with satisfactory early and midterm results. This technique could be an attractive alternative to conventional total arch replacement.

Total arch repair for acute type A aortic dissection with 2 modified techniques: open single-branched stent graft placement and reinforcement of the dissected arch vessel stump with stent graft.

BACKGROUND: In total arch replacement for patients with acute type A aortic dissection, anastomoses of the graft to the left subclavian artery and descending aorta are often difficult, and the arch vessel anastomosis is frequently performed at the site of dissection. To make this procedure easier and safer, we developed 2 modified techniques: open single-branched stent graft placement into the left subclavian artery and the descending aorta and reinforcement of the dissected arch vessel stump with a stent graft neointima. The feasibility and initial clinical results of these 2 new techniques are reported. METHODS AND RESULTS: Total arch repair with the 2 new techniques was performed in 26 patients with acute Stanford type A aortic dissection. Most patients had an uneventful postoperative course, and there were no surgical deaths. All implanted stent grafts were fully opened and not kinked, and there was no space or blood flow surrounding any of the stent grafts. The false lumen of the descending thoracic aorta closed with thrombus formation in 22 of 26 patients. Disappearance of the false lumen and recovery of the true lumen was observed in all of the dissected arch vessels. CONCLUSIONS: Open single-branched stent graft placement into the left subclavian artery and the descending aorta and reinforcement of the dissected arch vessel stump with a stent graft neointima are 2 simple and effective techniques that should make total arch repair an easier and safer procedure for acute type A aortic dissection.

[Modified total arch replacement plus stented elephant trunk implantation in acute type A aortic dissection: open single-branched stent graft placement].

OBJECTIVE: To explore the feasibility of open single-branched stent graft placement for simplified total arch replacement in acute Stanford type A aortic dissection. METHODS: Between June 2008 and September 2009, 23 patients with acute Stanford type A aortic dissection underwent total arch replacement plus open single-branched stent graft placement. When core nasopharyngeal temperature dropped to 68 H, the perfusion to lower body was discontinued. Left common carotid artery was transected at its ostium and its proximal stump closed. The arch was transected at a predetermined line between innominate artery and left common carotid artery. Through a transverse incision, the main stent graft of single-branched stent graft was inserted into true lumen of descending aorta. And the side arm stent graft was positioned into left subclavian artery. The transected stump of arch was reconstructed by inner proximal stent-free Dacron tube of main graft and outer Teflon felt. Subsequently continuous anastomosis was made to a 3-branched Dacron tube graft. RESULTS: Open single-branched stent graft placement was technically successful in all patients. The mean cardiopulmonary bypass time, aortic cross-clamp time, lower body arrest time and selective cerebral perfusion time were (161 ± 32), (97 ± 20), (21 ± 4) and (31 ± 6) min respectively. All patients were discharged from hospital without any complication. Their computed tomographic scans at 3 months postoperatively showed that all stent grafts were fully opened and not kinked. There was neither space nor blood flow surrounding the single-branched stent graft. The false lumen of descending aorta around stent graft closed with thrombus in all cases. CONCLUSION: Open single-branched stent graft placement is a new effective technique for simplified total arch replacement in acute type A aortic dissection. With this technique, the left subclavian artery anastomosis and distal aortic anastomosis at descending aorta can be avoided.

Open-branched stent graft placement makes total arch replacement easier for acute type a aortic dissection.

In total arch replacement for acute type A aortic dissection, the left subclavian artery anastomosis and the distal anastomosis at the descending aorta are often remarkably difficult because of the deep surgical field. We developed a single-branched stent graft that was constructed by adding a side arm stent-graft to a conventional aortic stent graft. By its open placement into the origin of the left subclavian artery and the descending aorta, the left subclavian artery anastomosis could be avoided, and the distal aortic anastomosis was performed at the proximal arch between the brachiocephalic artery and the left common carotid artery.

Cell cycle-related kinase supports ovarian carcinoma cell proliferation via regulation of cyclin D1 and is a predictor of outcome in patients with ovarian carcinoma.

Our previous study has suggested that the cell cycle-related kinase (CCRK) is a putative candidate oncogene in glioblastoma tumorigenesis. The potential oncogenic role of CCRK and its clinical/prognostic significance, however, in ovarian carcinoma are unclear. In this study, CCRK expression was examined by immunohistochemistry in a series of ovarian carcinoma tissues. Overexpression of CCRK was detected in 53% of the ovarian carcinomas, and it was positively correlated with an ascending histological grade and/or advanced clinical stage of the disease (p < 0.05). In addition, overexpression of CCRK in ovarian carcinoma was determined to be a strong and an independent predictor of short overall survival (p < 0.05). In ovarian carcinoma cells, CCRK knockdown by RNAi led to a G1 phase cell cycle arrest, while CCRK overexpression by stable transfection of CCRK-containing plasmid pcDNA-CCRK promoted cell proliferation in vitro and tumor growth in vivo. In addition, CCRK knockdown was found to reduce cyclin D1 expression. Consistently, CCRK overexpression increased cyclin D1 expression, and furthermore, a significant correlation between expression of CCRK and cyclin D1 in ovarian carcinomas was observed (p < 0.001). These findings suggest a potential important role of CCRK in the control of cell proliferation via regulation of cyclin D1 expression, and the overexpression of CCRK, as detected by immunohistochemistry, is an independent molecular marker for shortened survival time of patients with ovarian carcinoma.

[Effect of berberin hydrochloride on blood concentration of cyclosporine A in cardiac transplanted recipients].

OBJECTIVE: To observe the clinic effect of combined use of berberin hydrochloride (Ber) with cyclosporine A (CsA) on the blood concentration of CsA in heart transplanted recipients. METHODS: The blood concentration of CsA, liver-renal function and blood lipids in 22 heart transplanted recipients, who received Ber-CsA combined therapy, were measured. RESULTS: The whole blood steady state concentration of CsA, C0 and C2, in recipients after being treated with Ber-CsA significantly increased than those before applying Ber-CsA (P < 0.01), with the mean increment of 26% and 18% respectively; the dosage of CsA used decreased in 21 patients by 25-100 mg/d; and the Ber-CsA showed no significant effect on liver-renal function or blood lipids (P > 0.05). CONCLUSION: Combined use of CsA with Ber could markedly increase the blood concentration of CsA in heart transplanted recipients and reduce the dosage of CsA required, save the fee for medical service, and shows no obvious adverse reaction.

[Significance and mechanisms of overexpression of PAK1 gene in epithelial ovarian neoplasms].

OBJECTIVE: To investigate the significance and mechanisms of overexpression of p21-activated kinase 1 gene (PAK1) in epithelial ovarian neoplasms. METHODS: Immunohistochemistry, fluorescence in situ hybridization and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling methods were used to examine the protein expression and amplification of PAK1 and cell apoptosis in 30 benign ovarian adenomas, 20 borderline tumors and 80 ovarian carcinomas by tissue microarray. RESULTS: In immunohistochemistry study, overexpression of PAK1 protein was observed in 7 (25.9%) informative benign ovarian adenomas, 7 (36.8%) borderline tumors and 53 (68.8%) ovarian carcinomas. A significant inverse correlation of PAK1 overexpression and cell apoptosis was observed in these epithelial ovarian neoplasm cohorts (P = 0.002). In addition, 27/31 (87.1%) poorly differentiated (G3) carcinomas showed overexpression of PAK1, the frequency was significantly higher than that in tumors of G1 - G2 (26/46, 56.5% , P =0.01). In fluorescence in situ hybridization study, only 2 (4.7%) informative ovarian carcinomas showed amplification of PAK1 gene. None of the borderline and benign ovarian tumors showed PAK1 amplification. CONCLUSION: Overexpression of PAK1 protein may be involved in the tumorigenesis of epithelial ovarian neoplasms and it is associated closely with the malignant histological phenotype of ovarian carcinomas. Mechanism other than gene amplification of PAK1 may play a more important role in the regulation of protein expression of PAK1 in ovarian tumors.

[A preliminary proteomic analysis of tauopathies].

OBJECTIVE: To investigate the molecular mechanisms of tauopathies. Comparative proteomic analysis of brain proteins was employed to study 4 patients with tauopathies as compared with 4 controls. METHODS: The brains of subjects who died without clinical or pathological involvement of nervous system and brains of patients with tauopathies were obtained at autopsy. The brain proteins were run by immobilized pH gradient (IPG) isoelectric focusing electrophoresis as the first dimension, and then run by vertical SDS-PAGE as the second dimension. The maps were visualized by silver staining or colloidal coomassie blue and analyzed with Image Master 2D Elite software. The proteins of interest were in-gel digested and identified using MALDI-TOF mass spectrometry or MALDI-TOF/TOF tandem mass spectrometry. RESULTS: 18 protein spots were differentially expressed as compared with age-matched nondemented control brains which were identified as glyceraldehyde 3-phosphate dehydrogenase, uracil DNA glycosylase, human superoxide dismutase, isocitrate dehydrogenase subunit, synaptotagmin I, thioredoxin peroxidase 1, glial fibrillary acidic protein, p25 alpha, enoyl coenzyme A hydratase short chain 1, pyridoxine-5'-phosphate oxidase, Mn-superoxide dismutase and alpha enolase, antioxidant protein 2, ferritin heavy chain, glutamate dehydrogenase precursor, peptidyl-prolyl cis-trans isomerase A, serum albumin precursor and dihydropyrimidinase-related protein 2. CONCLUSIONS: We got a number of related-proteins of tauopathies. Some proteins are quite useful for discovering the molecular mechanisms of tauopathies and may be helpful for diagnosis and of treatment tauopathies.

[Proteomic comparison between human cerebellums and frontal lobes].

OBJECTIVE: To compare the results of proteomics between the cerebellum and frontal lobe of the aged. METHOD: Proteins were isolated from human cerebellums and frontal lobes and separated by two-dimensional (2D) gel electrophoresis. The proteins were then stained with silver or colloidal coomassie blue to produce a high-resolution map of the proteome. Selected proteins from this map were in-gel digested with trypsin and the resulting tryptic peptides were analyzed by MALDI-TOF mass spectrometry and MALDI TOF/TOF tandem mass spectrometry. The mass spectrometric data were used to identify the proteins through searches of the SWISSPROT protein sequence database. RESULTS: Three up-regulated protein spots in 2D gels of cerebellums were found as compared with those of frontal lobes. These protein spots were identified as antioxidant protein 2, creatine kinase precursor, fructose-bisphosphate aldolase C. Two down-regulated proteins in cerebellums were identified as pyruvate kinase M1 isozyme and glial fibrillary acidic protein. 30 common proteins, the expressions of which were not altered between cerebellums and frontal lobes, were also identified. CONCLUSIONS: These data will be used for future studies of differential protein expression in neurological disorders. Some proteins are useful for discovering the molecular mechanisms of degenerative dementia and brain aging.

[Effect of Ginkgo biloba extract on neuronal apoptosis in rabbit with kaolin-induced syringomyelia].

OBJECTIVE: To evaluate the effect of Ginkgo biloba extract (GBE) on neuronal apoptosis in rabbits with kaolin-induced syringomyelia. METHODS: Twenty-four of 30 Chinese white rabbits were subjected to injection of 25% kaolin mixed with equal volume (0.6 ml) of cerebrospinal fluid drawn from the cisterna magna under ketamine anesthesia. Twelve of these 24 rabbits then received intravenous injection of 5 ml of GBE (5 ml/days for 14 days, GBE treatment group) while the other 12 were treated with the same amount of saline administered in similar manner (saline group). The 6 rabbits without kaolin treatment received a sham operation to serve as the control group. At different time points after the operation, the rabbits were killed and the spinal cord samples examined by immunohistochemistry. RESULTS: Histologically, ischemia and edema in the cervical cord of rabbits in GBE treatment group were less severe than those in saline group. TUNEL-positive and bax-positive neurons were less numerous in GBE treatment group than in saline group, and the former group showed more Bcl-2-positive neurons. The number of apoptotic neurons reached the peak level on day 7 after kaolin injection. CONCLUSION: GBE can ameliorate kaolin-induced hydrocephalus in the upper cervical cord and inhibit kaolin-induced neuron apoptosis.

[Two-dimensional electrophoregram for proteomic analysis of rat brain with intrahippocampal amyloid beta injection and normal rat brain].

OBJECTIVE: To investigate the molecular mechanisms of Alzheimer's disease by comparing global protein patterns in two-dimensional electrophoregram (2-DE) of the brain of rats with intrahippocampal amyloid beta injection and normal rats. METHODS: From adult SD rats with intrahippocampal injection of amyloid beta, 200 microg brain proteins were extracted with 9 mol/L urea, 4% CHAPS, 1% DTT, 0.5% CA and a cocktail of protease inhibitors. Immobilized pH gradient (IPG) isoelectric focusing electrophoresis of the extracted proteins was performed to obtain the electrophoretogram of the first dimension, with the second dimension obtained by vertical SDS-PAGE. The electrophoretograms were visualized using silver staining and analyzed with ImageMaster 2D-Elite software. RESULTS: On average, 496 and 491 protein spots could be obtained in the electrophoregraphs for rats with amyloid beta and the control rats, respectively, and 30 of these spots exhibited quantitative changes. Another 11 and 6 spots were exclusively shown on the protein maps for amyloid beta-treated rats and control rats, respectively. CONCLUSION: The differentially displayed proteins in the brain identified between the rats with intrahippocampal amyloid beta injection and control rats may provide further insight into the pathogenesis of Alzheimer's disease and useful clues for developing new drugs for its treatment.