NCAPG is a prognostic biomarker associated with vascular invasion in hepatocellular carcinoma.

OBJECTIVE: Vascular invasion is closely associated with tumor recurrence and poor patient outcomes in individuals diagnosed with hepatocellular carcinoma (HCC). In this study, we explored the potential value of NCAPG as a prognostic biomarker of vascular invasion in HCC patients. MATERIALS AND METHODS: Two Gene Expression Omnibus (GEO) datasets (GSE14520 - GPL3721 Subset; GSE67140 - GPL8786) were utilized to explore the relationship between genes and HCC-associated vascular invasion. Hub genes associated with vascular invasion were identified through analyses of Cytoscape using the Cytohubba plugin, and relationships between specific genes and patient survival outcomes were assessed through univariate and multivariate analyses of the TCGA-Liver Hepatocellular Carcinoma (TCGA-LIHC) database. RESULTS: In total, 10 hub genes were associated with vascular invasion in the two analyzed GEO datasets. Importantly, non-SMC condensin I complex subunit G (NCAPG) overexpression was correlated with poor prognosis for patients in the TCGA-LIHC database. NCAPG was identified as an independent predictor of HCC patient overall survival (OS) (HR 2.543; 95% CI 1.224, 5.285; p = 0.012) and disease-free survival (DFS) (HR 2.034; 95% CI 1.160, 3.566; p = 0.013) in a multivariate Cox analysis. NCAPG expression status, vascular invasion status, tumor status, and AJCC T stage were independent predictors of OS, with a concordance index (c-index) value of 0.713 (95% CI, 0.671, 0.756). NCAPG expression levels were related to hypomethylation status and were positively correlated with tumor immune cell infiltration and immune cell-related biomarker expression. CONCLUSIONS: NCAPG upregulation is associated with poor prognosis in hepatocellular carcinoma patients with vascular invasion.

High HBXIP expression is related to poor prognosis in HCC by extensive database interrogation.

OBJECTIVE: The involvement of HBXIP in cancer development and cancer cell survival is well known. This work probed the potential of HBXIP as a prognostic biomarker in hepatic cell cancer (HCC). MATERIALS AND METHODS: First, pan-cancer analysis of HBXIP expression was conducted using The Cancer Genome Atlas (TCGA) database to validate the expression of HBXIP in different cancers. The GSE14520 (GPL3721 Subset) database was used to validate HBXIP in HCC. The association between survival outcomes and prognostic factors was assessed employing univariate and multivariate survival analyses for TCGA Liver Hepatocellular Carcinoma. The biological function of the HBXIP Gene was annotated by gene set enrichment analysis. The relationship between HBXIP expression and immune cells and immune markers was analyzed from the Gene Expression Profiling Interactive Analysis (GEPIA) database. RESULTS: Malignant tissues demonstrated evident upregulation of HBXIP at transcriptional and protein levels over normal tissues (p < 0.05) with this elevated expression linked to an advanced tumor stage in HCC cohorts. Univariate analysis revealed an evident correlation emerged between prognosis and HBXIP for GSE14520 databases (p < 0.05). The disease-free survival (DFS) and overall survival (OS) (five-year values) were lower in samples demonstrating elevated HBXIP (HR: 2.413; 95% CI 1.601, 3.638; p < 0.001) and (HR: 1.613; 95% CI 1.446, 1.844; p = 0.003), respectively vs. lower HBXIP expression. HBXIP emerged as an independent factor in OS prognosis (HR 2.184; 95% CI 1.495, 3.196; p < 0.001) and DFS (HR 1.764; 95% CI 1.261, 2.466; p < 0.001), respectively according to multivariate analysis. Further, multiple Cox analyses in the validation cohort revealed that independent factors for OS were HBXIP, AJCC T stage, vascular invasion, and tumor status with the C-index score of 0.727 (95% CI, 0.704 to 0.750). HBXIP level showed a significantly positive association with tumor immune cell infiltration, and biomarkers of immune cells; besides, the rectum Rho GTPase effectors signaling pathway was also identified. CONCLUSIONS: HCC advancement and survival involves HBXIP, which also emerged as a functional biomarker for HCC survival prediction.

Isolation and identification of phase I metabolites of resibufogenin in rats.

1. Resibufogenin (1), a major bufadienolide of Chinese medicine Chan Su, had a wide range of pharmacological activities. In present work, the metabolism of 1 in male Sprague-Dawley rats was investigated by identifying the metabolites of resibufogenin excreted in rat bile. 2. Following an oral dose of 60 mg/kg resibufagenin, nine metabolites were isolated from bile of rats, and their structures were identified as 3-keto- resibufogenin (2), 3-epi-resibufogenin (3), 5ß-hydroxy-3-epi-resibufogenin (4), 1α, 5ß-dihydroxy-3-epi-resibufogenin (5), 3α, 5ß, 14α, 15ß-tetrahydroxyl-bufa- 20, 22-dienolide (6), 3α, 14α, 15ß-trihydroxy-bufa-20, 22-dienolide (7), 3-epi- 5ß-hydroxy-bufalin (8), 12α, 16ß-dihydroxy-3-epi-resibufogenin (9), and 5ß, 16ß-dihydroxy-3-epi-resibufogenin (10), respectively, on the basis of widely spectroscopic methods including 2D-NMR technology. It is first time to describe the metabolites of 1 in vivo, and metabolites 5-7 and 9-10 are novel. 3. On the basis of these identified metabolites, a possible metabolism pathway for 1 in rats has been proposed. This is the first systematic study on the phase I metabolites of resibufogenin.

Dopamine receptors set the pattern of activity generated in subthalamic neurons.

Information processing in the brain requires adequate background neuronal activity. As Parkinson's disease progresses, patients typically become akinetic; the death of dopaminergic neurons leads to a dopamine-depleted state, which disrupts information processing related to movement in a brain area called the basal ganglia. Using agonists of dopamine receptors in the D1 and D2 families on rat brain slices, we show that dopamine receptors in these two families govern the firing pattern of neurons in the subthalamic nucleus, a crucial part of the basal ganglia. We propose a conceptual frame, based on specific properties of dopamine receptors, to account for the dominance of different background firing patterns in normal and dopamine-depleted states.

[Inhibitive effect of puerarin on increased NO production by neonatal cardiomyocytes during hypoxia/reoxygenation injury].

OBJECTIVE: To investigate the effect of puerarin (Pue) on nitric oxide (NO) produced by neonatal rat cardiomyocytes during hypoxia/reoxygenation injury. METHOD: NO contents in the culture supernatants sampled from different groups (control, model, and therapeutic groups with 1, 0.1, 0.01 g.L-1 Pue) at different time were assayed with nitrate reductase method. RESULT: NO content of model increased after reperfusion (P < 0.01), while it increased sharply at 6 h after reperfusion and kept thereafter. Only at 6 h after reperfusion did Pue in these three doses inhibit NO production (P < 0.01) and kept to the 24 h after reperfusion. CONCLUSION: Pue exerts inhibitive effect only after NO production is enhanced sharply during hypoxia/reperfusion injury in a dose-dependent trend.

Differential effects of L-deprenyl on MPP+- and MPTP-induced dopamine overflow in microdialysates of striatum and nucleus accumbens.

The present studies investigated the effects of L-deprenyl, 1-methyl-4-phenylpyridinium ion (MPP+) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on the efflux of dopamine and its metabolites in microdialysates of striatum and nucleus accumbens in rats. L-Deprenyl or L-amphetamine perfusion into striatum had no effects on basal dopamine efflux, though L-deprenyl reduced the basal efflux of dihydroxyphenylacetic acid and homovanillic acid. MPP+ or MPTP perfusion into striatum significantly increased the dopamine efflux, and the action of MPTP was more potent than that of MPP+. Pretreatment with L-deprenyl antagonized the actions of MPP+ and MPTP. The striatal dopamine efflux of rats was gradually restored by itself after the overflow caused by 2-h perfusion of the dopaminergic neurotoxins, while L-deprenyl could not accelerate the recovery. Perfusion with L-deprenyl or L-amphetamine, but not pargyline, into nucleus accumbens increased the dopamine efflux in a dose-dependent fashion, which could be antagonized by haloperidol pretreatment. MPP+ or MPTP perfusion into nucleus accumbens also increased the dopamine efflux, and the action of MPTP was also more potent than that of MPP+. Pretreatment with L-deprenyl could not antagonize the actions of MPP+ and MPTP. These findings suggest that L-deprenyl, MPP+ and MPTP induce differential effects on nigrostriatal and mesolimbic dopaminergic pathways in vivo. L-Deprenyl has neuroprotective rather than neurorestorative action against MPP+- and MPTP-induced dopamine overflow from striatum. Further, L-deprenyl-induced dopamine overflow from nucleus accumbens may explain the amphetamine-like reinforcing property of L-deprenyl.

Modulation of medical prefrontal cortical D1 receptors on the excitatory firing activity of nucleus accumbens neurons elicited by (-)-Stepholidine.

(-)-Stepholidine (SPD), with D1 agonistic action, elicited an excitatory firing activity of nucleus accumbens (NAc) neurons by intravenous administration, but this effect was hardly observed by iontophoresis of SPD into the NAc. The present study intends to determine whether D1 receptors in the medial prefrontal cortex (mPFC) are involved in the action of SPD on the firing activity of NAc neurons in the chloral hydrate-anesthetized male rats. The results showed that the intra-mPFC microinjected SCH-23390 (D1 antagonist, 30 mM), but not the D2 antagonist spiperone (30 mM), significantly attenuated the enhanced firing activity induced by intravenous injection of SPD (2 mg/kg). Similarly, the excitatory firing of NAc neurons was also exhibited by the microinjection of either SPD or D1 agonist SKF-38393 into the mPFC. The SPD-induced excitatory effect was in a dose-dependent way from 277.8 +/- 51.3% (10 mM) to 1105.4 +/- 283.5% (30 mM) of NAc basal firing, which was completely reversed by SCH-23390 (i.v.). Furthermore, the direct D1 agonistic action of SPD on the mPFC neuron was observed with microiontophoresis. These results indicate that SPD possesses a direct agonistic action on the mPFC D1 receptors, by which it modulates the firing activity of NAc neurons.

[Radiosensitivity of different ploidy pollen in poplar and its application].

Through investigation on germinating rate and process of unreduced diploidy pollen and monoploid pollen in Populus tomentosa x P. alba and P. tomentosa, it was proved that there were different radiosensitivities to 60Co gamma-ray among different ploidy pollen, i.e. the radiosensitivity of monoploidy pollen was two times higher than diploidy pollen. To overcome the problems of late germination and poor compatibility of diploidy pollen during fertilization, a certain dose of radiation can be applied to stimulating the diploidy pollen germination, meanwhile to restrain of kill some monoploidy pollen based on their different radiosensibilities. Therefore the rate of germination and faster germinating process of diploidy pollen on stigmas can be increased relatively to certain extent. The efficiency of radiation was proved by breeding practice. In the hybidization of (P. alba x P. glandulosa) x (P. tomentosa x P. bolleana), 3.8% of triploid could be produced by 1,470 rad treatment; while no triploid is examined in the control. 12.9% of triploids were produced by the treatment of 1,680 rad from the above cross.

Electrophysiological study on biphasic firing activity elicited by D(1) agonistic-D(2) antagonistic action of (-)-stepholidine in nucleus accumbens.

Our previous work has demonstrated that (-)-stepholidine (SPD) has dual action, ie D(1) agonistic-D(2) antagonistic action on DA receptors in the nigra-striatal dopamine (DA) system. The present study attempted to ascertain its dual action on the mesolimbic DA system. The firing activities of the nucleus accumbens (NAc) neurons were extracellularly recorded with intravenous and iontophoretic administration of the drug in 6-hydroxydopamine (6-OHDA)-lesioned and intact rats. The results showed that SPD produced a consistently biphasic firing of NAc neurons during the cumulative doses of 0.02 2 mg/kg, iv. When the rats were pretreated with D(2) antagonist spiperone, SPD only exerted an increasing effect, which was subsequently reversed by the D(1) antagonist SCH-23390. Moreover, SCH-23390 could prevent the rate of increase elicited by SPD at high doses, presumably due to the D(1) agonistic action of SPD on the activity of NAc neuron. On the other hand, the inhibition of NAc firing elicited by either D(2) agonist LY171555 or D(1)/D(2) agonists apomorphine was completely reversed by SPD, suggesting an antagonistic action of SPD to D(2) receptors. In 6-OHDA-lesioned rats, iontophoresis of SPD also had an inhibitory effect in the majority of NAc neurons (91%) as SKF-38393 did. This inhibition could be completely blocked by the ejection of SCH-23390, but not by spiperone. These results indicate that SPD also has a D(1) agonistic-D(2) antagonistic dual action on NAc neuron activity, which may be beneficial to the treatment of schizophrenia.

Behavioral characteristics of olanzapine: an atypical neuroleptic.

AIM: To assess the atypical neuroleptic properties of a novel antipsychotic agent, olanzapine (Ola). METHODS: The action of Ola on apomorpine (Apo)-induced climbing behavior, 5-hydroxy-dl-tryptophan (5-HTP)-induced head twitch response, oxotremorine-induced tremor, and the conditioned avoidance behavior in mice were observed. The catalepsy of mice induced by Ola was also investigated. The single unit extracellular recording technique was used to compare the spontaneous firing rate changes of dopamine (DA) cells in the ventral tegmental area (VTA, A10) and the substantia nigra pars compact (SNC, A9) in rats after i.v. Ola. RESULTS: Ola antagonized the climbing behavior (ED50 1.8 mg.kg-1, p.o.), head twitch behavior (ED50 0.3 mg.kg-1, p.o.), and tremor (ED50 5.2 mg.kg-1, p.o.) in mice. In a conditioned avoidance paradigm in mice, Ola inhibited the avoidance response with an ED50 of 2.72 mg.kg-1 (p.o.). However, the catalepsy was not induced by Ola in mice even under a very high dose of 100 mg.kg-1 (p.o.). Ola selectively increased the firing rate of DA cells in the VTA, but failed to affect that of SNC DA cells. CONCLUSION: Ola distinguished itself from the typical neuroleptic (e.g. haloperidol, Hal) and took resemblance of the atypical neuroleptic (e.g. clozapine, Clo) in 3 aspects: 1) the multiple receptor pharmacodynamics involving D1/D2, 5-HT2 and M-ACh receptors; 2) dose-response separation between the block of conditioned avoidance response and catalepsy induction; and 3) the specificity of action sites of firing rates upon acute drug challenge.

I-stepholidine facilitates inhibition of mPFC DA receptors on subcortical NAc DA release.

AIM: To determine whether the D1 agonistic action of (-)-stepholidine (SPD) on the medial prefrontal cortex (mPFC) neuron is involved in the modulation of evoked subcortical dopamine (DA) release from nucleus accumbens (NAc) of rats. METHODS: With the microinjection of SPD into the mPFC, the ventral tegmental area (VTA)-stimulated or amphetamine (AMP)-evoked DA efflux in the NAc was detected by microdialysis + HPLC-ECD in the 6-hydroxydopamine (6-OHDA)-lesioned and intact rats. RESULTS: The depletion of DA in the mPFC did not modify both the basal level and the VTA-stimulated DA efflux in the NAc, but significantly facilitated the AMP (20 mumol.L-1)-evoked DA efflux within the NAc. It indicates that the mPFC DA system is involved in the regulation of evoked DA release in the NAc. Besides, the AMP-evoked increase of the extracellular DA release in the NAc was significantly attenuated by SPD (10, 30 mmol.L-1) microinjection into the mPFC, though this injection of SPD could not alter the response of DA release by the stimulation of the VTA. Furthermore, the inhibitory effect of SPD on the AMP-evoked DA efflux could be partially reversed by intravenous administration of D1 antagonist Sch-23390 (1 mg.kg-1), but not by D2 antagonist spiperone. CONCLUSION: SPD is capable of enhancing the function of D1 receptors in the mPFC, by which it facilitates the inhibition of DA release in the NAc.

[Destruction of NA innervation in the rat hippocampus resists the hypoxia-induced corticosterone secretion].

With microinjection of 6-hydroxydopamine (6-OHDA) into the ventral hippocampus (VHIP) of rats, the effect of 6-OHDA lesion of hippocampal noradrenergic system on the plasma corticosterone (CS) response to acute hypoxia was investigated. It was found that the plasma CS response to inhalation of 10.4% O2 (30 min) was increased markedly; while the plasma CS level and the noradrenaline (NA) contents in hippocampus were decreased (-33.2% and -38.5%, respectively) 2 weeks after microinfusion of 6-OHDA (8 micrograms/2 microliters) into VHIP; however, the CS response to acute hypoxia was unaffected by 6-OHDA lesion of VHIP. The results above suggest that NA in hippocampus may participate in the regulation of the hypoxia-induced increase of plasma CS concentration in rats.

Comparison of (-)-stepholidine and D1 or D2 agonists on unit firing of globus pallidus in 6-hydroxydopamine-lesioned rats.

(-)-Stepholidine(SPD), isolated from the Chinese herb Stephania, is demonstrated to be a DA antagonist, but it also shows D1 agonistic action on rotational behavior in unilateral 6-hydroxydopamine (6-OHDA) lesioned rats. The present study further ascertains its D1 agonistic property on firing activity of globus pallidus (GP) in control and 6-OHDA-lesioned rats. In the control rats, the firing activities of the GP neurons elicited by DA agonists (i.v.), such as apomorphine (D1/D2), SKF38393 (D1), and LY171555 (D2), were readily reversed by SPD (4 mg/kg, i.v.); but SPD, per se, induced variable alterations. In the 6-OHDA-lesioned rats, apomorphine, SKF38393 and LY171555 displayed the marked inhibition as well as excitation on the unit firing. The individual firing variations (87.1+/-17.8, 55.1+/-15.7 and 62.1+/-16.7%, respectively) were much larger than those in the control group, and were completely abolished by SPD (2 mg/kg). However, SPD also showed D1 partial agonistic action on the GP neuron firing. Moreover, the pre-blockade of D2 receptors with spiperone (0.5 mg/kg, i.v.), SPD exhibited the D1 agonist action which was reversed by the D1 antagonist SCH23390. These results suggest that SPD has a dual action on the GP neuron firing in the 6-OHDA-lesioned rats, i.e., antagonist to D2 DA receptors and partial agonist to D1 receptors.

Effect of desipramine on spontaneous activity of hippocampal CA1 neuron after transient cerebral ischemia in rats.

AIM: To study the spontaneous firing of CA1 neurons in rat hippocampus after transient cerebral ischemia and the effect of desipramine (Des) on the post-ischemic electric activity of CA1 neurons. METHODS: Single-unit extracellular recordings were performed in rats on d 3 after 10 min of cerebral ischemia by occlusion of 4 arteries. Des and saline were injected into a tail vein. The histological changes of CA1 neurons was assessed by the neuronal density of the CA1 sector. RESULTS: The spontaneous firing rate of CA1 neurons on d 3 after ischemia was enhanced in comparison with the control value. Des (0.2 and 0.4 mg.kg-1, i.v., n = 5 & 6, respectively) reduced dose-dependently the increase of firing rate with maximal inhibition by 6 min (58% & 85%) to 9 min (69% & 94%) (vs vehicle group, P < 0.01). About 50% cells in CA1 region showed necrotic changes. CONCLUSION: Des antagonized the hyperexcitability of CA1 neurons after cerebral ischemia.

[Participation of septum on the increase of plasma cortisol level induced by stimulation of the greater splanchnic nerve of cats].

Experimens were performed on 36 cats, showing that stimulation of the central end of the left major splanchnic nerve induced an increase of the plasma cortisol level. The centripetal effect was abolished by lesioning the septal nuclei, just as the same as injection of propranolol into the nuclei; but phentolamine had no such preventive action. All the findings show that it is the noradrenergic beta-receptor system of the septal nuclei that exerts a considerable effect on cortisol secretion.

Capillary gas chromatographic analysis of carbohydrates of Legionella pneumophila and other members of the family Legionellaceae.

Legionella pneumophila, the causative agent of Legionnaires disease, and related organisms have previously been characterized primarily by conventional bacteriological methods, DNA-DNA hybridization, antigenic analysis, and fatty acid analysis. By capillary gas chromatographic analysis for carbohydrates, we have shown that muramic acid and glucosamine, characteristic markers of bacterial cell walls, were present in samples of L. pneumophila and a group of legionella-like organisms. Some bacterial samples contained two unusual isomeric aminodideoxyhexoses (X1 and X2). L. pneumophila was characterized by the absence of fucose and the presence of the peak X1. Tatlockia micdadei (Legionella micdadei) was distinguishable by the presence of large amounts of rhamnose and fucose and by the absence of X1 and X2. Fluoribacter strains were much more variable in their carbohydrate composition. These data suggest that, in addition to other reported techniques, carbohydrate profiling by capillary gas chromatography can be a valuable diagnostic method in reference microbiology laboratories for differentiating members of the family Legionellaceae.

Capillary gas chromatographic analysis of alditol acetates of neutral and amino sugars in bacterial cell walls.

Several improvements in the preparation of alditol acetates of neutral and amino sugars and in the preparation of glass capillary columns for their separation are described. Modifications in sample preparation permitted the simultaneous processing of multiple samples and eliminated extraneous background peaks. Efficient and inert columns were tailor-made for the separation of alditol acetates of neutral and amino sugars by leaching glass capillaries with aqueous hydrochloric acid and dynamically coating with SP-2330.