Exploration in the Therapeutic and Multi-Target Mechanism of Ketamine on Cerebral Ischemia Based on Network Pharmacology and Molecular Docking.

Background: Ketamine is famous for its dissociative anesthetic properties. It is also analgesic, anti-inflammatory and anti-depressant, and even has a cerebral protective effect. We searched the evidence of the correlation between ketamine target and clinical efficacy and utilized network pharmacology to gather information about the multi-target mechanism of ketamine against cerebral ischemia (CI). We found that ketamine's clinical significance may be more extensive than previously thought. Methods: The drug target of ketamine and CI-related genes were predicted by SwissTargetPrediction, DrugBank, PubChem, GeneCards and DisGeNET databases. The intersection of ketamine's drug-targets and CI-related genes was analyzed by using GO and KEGG. We predicted the molecular docking between the potential target and ketamine. Results: The results indicated that the effect of ketamine on CI was primarily associated with the target of α-synuclein (SNCA), muscarinic acetylcholine receptor M1 (CHRM1) and nitric oxide synthase 1 (NOS1). It principally regulates the signal pathways of circadian transmission, calcium signaling pathway, dopaminergic synapse, cholinergic synapse and glutamatergic synapse. Molecular docking analysis exhibited that hydrogen bond and Pi-Pi interaction were the predominant modes of interaction. Conclusion: There are protein targets affected by ketamine in the treatment of CI. Three pivotal targets involving 298 proteins, SNCA, CHRM1 and NOS1, have emerged as multi-target mechanisms for ketamine in CI therapy. Similarly, this study also provides a new idea for introducing network pharmacology into the evaluation of multi-targeted drugs for CI and cerebral protection.

Biliverdin administration regulates the microRNA-mRNA expressional network associated with neuroprotection in cerebral ischemia reperfusion injury in rats.

Inflammatory response has an important role in the outcome of cerebral ischemia reperfusion injury (CIR). Biliverdin (BV) administration can relieve CIR in rats, but the mechanism remains unknown. The aim of the present study was to explore the expressional network of microRNA (miRNA)­mRNA in CIR rats following BV administration. A rat middle cerebral artery occlusion model with BV treatment was established. After neurobehavior was evaluated by neurological severity scores (NSS), miRNA and mRNA expressional profiles were analyzed by microarray technology from the cerebral cortex subjected to ischemia and BV administration. Then, bioinformatics prediction was used to screen the correlation between miRNA and mRNA, and 20 candidate miRNAs and 33 candidate mRNAs were verified by reverse transcription­quantitative polymerase chain reaction. Furthermore, the regulation relationship between ETS proto­oncogene 1 (Ets1) and miRNA204­5p was examined by luciferase assay. A total of 86 miRNAs were differentially expressed in the BV group compared with the other groups. A total of 10 miRNAs and 26 candidate genes were identified as a core 'microRNA­mRNA' regulatory network that was linked with the functional improvement of BV administration in CIR rats. Lastly, the luciferase assay results confirmed that miRNA204­5p directly targeted Ets1. The present findings suggest that BV administration may regulate multiple miRNAs and mRNAs to improve neurobehavior in CIR rats, by influencing cell proliferation, apoptosis, maintaining ATP homeostasis, and angiogenesis.

Biliverdin administration ameliorates cerebral ischemia reperfusion injury in rats and is associated with proinflammatory factor downregulation.

Biliverdin (BV), one of the heme oxygenase-1 (HO-1) catalytic products, has been demonstrated to have protective effects in liver ischemia reperfusion injury (IRI). The present study aimed to explore the effects of BV on cerebral IRI, and to investigate the potential mechanisms thereof. Adult male SD rats, weighing 200-240 g, were randomly divided into sham (group S), cerebral ischemia reperfusion control (group C) and BV (group BV) groups. Rats in group C underwent transient middle cerebral artery occlusion (tMCAO) and received 2 ml normal saline; rats in group BV received BV (35 mg/kg) intraperitoneally 15 min prior to reperfusion and 4 h after reperfusion, then twice a day thereafter for 5 days. Group S served as the control. Neurological Severity Scores (NSS) were evaluated at days 1-5 following reperfusion. Staining with 2, 3, 5-triphenyltetrazolium chloride was performed to determine the cerebral infarction at 48 h post reperfusion. mRNA expression levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1ß, inducible nitric oxide synthase (iNOS) and HO-1 in the ischemic cerebral cortex were detected via reverse transcription-quantitative polymerase chain reaction at 3, 6, 12 and 24 h after reperfusion. Western blotting was used to detect the protein expression levels at 3 h after reperfusion. Compared with group S, the NSS, cerebral infarct volume, and the mRNA and protein expression levels of TNF-α, IL-6, IL-1ß, iNOS and HO-1 of Group C were significantly increased (P<0.05). However, BV administration significantly improved and reduced these expression levels (P<0.01). The present study indicates that BV is able to ameliorate cerebral IRI in rats and that the mechanism may be associated with the downregulation of proinflammatory factors.

Fine-needle aspiration with measurement of parathyroid hormone levels in thyroidectomy / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>Hypoparathyroidism is one of the most serious complications of thyroidectomy. It is important to identify the parathyroid glands during thyroidectomy. In order to find an economic, simple and less traumatic way to identify the parathyroid glands and testify its feasibility, fine-needle aspiration of suspected parathyroid tissue was used to measure the parathyroid hormone (PTH) levels during the surgical procedure.</p><p><b>METHODS</b>From Nov. 2011 to Apr. 2012, 50 patients were recruited for thyroid surgery in the Sun Yat-sen University Cancer Centre. During surgery, fine-needle aspiration of suspected tissues, including parathyroid gland, thyroid gland, muscle, fat tissue, and lymph node, was performed, the PTH levels were measured. In addition, the tissues above-mentioned were taken to pathological examination. Statistical processing was adopted to determine the sensitivity and specificity of intraoperative fine-needle aspiration with measurement of PTH level in finding the pathology of the parathyroid gland.</p><p><b>RESULTS</b>There were 237 tissues from 50 patients in total, and 45 of them were certified as the parathyroid glands by pathology. Intra-operative PTH (ioPTH) of the tissues in forty-four cases were higher than 600 ng/L, ioPTH of the tissues in one case was lower than 600 ng/L, and it was 160 ng/L. The highest ioPTH in other cases was 537.7 ng/L. The sensitivity was 97.8%. The specificity was 100%. The difference between the sensitivity and the specificity of two groups was not statistically significant, and P > 0.05. The level of PTH of parathyroid gland were much higher than other tissues, and P < 0.001.</p><p><b>CONCLUSIONS</b>The level of ioPTH of parathyroid gland were far higher than thyroid, muscle, fat, lymph node. It is an economic, fast and less traumatic way to identify the parathyroid gland by using the fine-needle aspiration of the parathyroid tissue with measurement of PTH levels. The sensitivity and the specificity are high. It can be used in the thyroidectomy to identify the parathyroid glands.</p>

Deciphering the effects of gene deletion on yeast longevity using network and machine learning approaches.

Longevity is one of the most basic and one of the most essential properties of all living organisms. Identification of genes that regulate longevity would increase understanding of the mechanisms of aging, so as to help facilitate anti-aging intervention and extend the life span. In this study, based on the network features and the biochemical/physicochemical features of the deletion network and deletion genes, as well as their functional features, a two-layer model was developed for predicting the deletion effects on yeast longevity. The first stage of our prediction approach was to identify whether the deletion of one gene would change the life span of yeast; if it did, the second stage of our procedure would automatically proceed to predict whether the deletion of one gene would increase or decrease the life span. It was observed by analyzing the predicted results that the functional features (such as mitochondrial function and chromatin silencing), the network features (such as the edge density and edge weight density of the deletion network), and the local centrality of deletion gene, would have important impact for predicting the deletion effects on longevity. It is anticipated that our model may become a useful tool for studying longevity from the angle of genes and networks. Moreover, it has not escaped our notice that, after some modification, the current model can also be used to study many other phenotype prediction problems from the angle of systems biology.

H2S protects hippocampal neurons from anoxia-reoxygenation through cAMP-mediated PI3K/Akt/p70S6K cell-survival signaling pathways.

The study aims to investigate the effect of hydrogen sulfide (H(2)S) on the phosphatidylinositol 3-kinase (PI3K)/Akt/p70 ribosomal S6 kinase (p70S6K) signal transduction pathway after oxygen glucose deprivation/reoxygenation (OGD/R) in the rat hippocampus. Newborn Wister rats were decapitated under anesthesia, and hippocampal tissue was dissected. Cells were plated at 1.0 × 10(5) cells/mL on polylysine-treated 96-well and 6-well plates. After 7 days in culture, cells were randomly assigned to six groups: control, OGD/R, sodium hydrosulfide (NaHS) following OGD/R, NaHS/triciribine following OGD/R, NaHS/rapamycin following OGD/R, and NaHS/triciribine/rapamycin following OGD/R. Neuronal purity and cell viability were assessed in each group, as well as apoptosis and expression of cyclic adenosine 3', 5'-monophosphate (cAMP), PI3K, Akt, and p70S6K. NaHS enhanced cAMP concentration and expression of PI3K, Akt, and p70S6K. In addition, neuronal viability was increased and apoptotic neuronal numbers decreased (P<0.01). Triciribine inhibited Akt and p70S6K, as well as decreased cell survival and viability compared with the NaHS group (P<0.05 or P<0.01). Rapamycin resulted in decreased p70S6K expression and neuronal viability, as well as increased number of apoptotic neurons compared with the NaHS group (P<0.05 or P<0.01). H(2)S acted via cAMP-mediated PI3K/Akt/p70S6K signal transduction pathways to inhibit hippocampal neuronal apoptosis and protect neurons from OGD/R-induced injury.

[Interaction between endogenous cystathionine synthase/hydrogen sulfide and heme oxygenase-1/carbon monoxide systems during myocardial ischemic-reperfusion: experiment with rats].

OBJECTIVE: To investigate the interaction between the cystathionine synthase gene (CBS)/hydrogen sulfide (H(2)S) and heme oxygenase-1 (HO-1)/carbon oxide (CO) systems during myocardial ischemia-reperfusion. METHODS: Twenty-four SD rats were randomly divided into 4 equal groups: ischemia-reperfusion (I/R) group undergoing intraperitoneal injection of normal saline (NS) and then 30 min later undergoing ligation of the left coronary artery (LCA) for 30 min, ischemia-reperfusion + zinc protoporphyrin (I/R + Z) group undergoing intraperitoneal injection of 45 micromol/kg zinc protoporphyrin, a HO-1 inhibitor, 30 min before the ligation of LCA, ischemia-reperfusion + hydroxylamine (I/R + H) group, undergoing intraperitoneal injection of 5 mmol/L hydroxylamine, a CBS inhibitor, 30 min before the ligation of LCA, and ischemia-reperfusion + zinc protoporphyrin + hydroxylamine (I/R + Z + H) group undergoing intraperitoneal injection of zinc protoporphyrin and hydroxylamine 30 min before the ligation of LCA. Another 6 rats underwent sham operation and intraperitoneal injection of NS 30 min before the operation and were used as control (C) group. The rats were killed 2 h after the reperfusion with their hearts taken out. Electron microscope was used to observe the mitochondrial structure. Homogenate of myocardium was prepared to examine the concentrations of H(2)S, CO, glutathione (GSH), malonyldialdehyde (MDA), activity of superoxide dismutase (SOD), and expression level of CBS-mRNA and HO-1-mRNA. RESULT: The concentration of CO, H(2)S, GSH, and MDA increased, SOD activity decreased, and HO-1-mRNA and CBS-mRNA expression levels increased in Group I/R as compared with Group C (all P < 0.01). In Group I/R + Z the CO level decreased, H(2)S and GSH levels increased, and HO-1-mRNA expression level decreased, but CBS-mRNA expression level increased as compared with Group I/R (all P < 0.05). In Group I/R + H the H(2)S and GSH levels decreased, CO level increased, and HO-1-mRNA expression level increased and CBS-mRNA expression level decreased as compared with Group I/R (all P < 0.05). In Group I/R + Z + H the H(2)S, CO, GSH, and SOD levels decreased, MDA level increased, and HO-1-mRNA and CBS-mRNA expression levels decreased as compared with Group I/R (all P < 0.05). Electron microscopy found that mitochondrial structure was destroyed in the Groups I/R, I/R + Z, I/R + H, and most seriously in Group I/R + Z + H. CONCLUSION: Both CBS/H(2)S and HO-1/CO systems play a protective role in myocardial ischemia-reperfusion and they interact on each other.