Differential gene expression profiles between two subtypes of ischemic stroke with blood stasis syndromes.

Ischemic stroke is a cerebrovascular thrombotic disease with high morbidity and mortality. Qi deficiency blood stasis (QDBS) and Yin deficiency blood stasis (YDBS) are the two major subtypes of ischemic stroke according to the theories of traditional Chinese medicine. This study was conducted to distinguish these two syndromes at transcriptomics level and explore the underlying mechanisms. Male rats were randomly divided into three groups: sham group, QDBS/MCAO group and YDBS/MCAO group. Morphological changes were assessed after 24 h of reperfusion. Microarray analysis with circulating mRNA was then performed to identify differential gene expression profile, gene ontology and pathway enrichment analyses were carried out to predict the gene function, gene co-expression and pathway networks were constructed to identify the hub biomarkers, which were further validated by western blotting and Tunel staining analysis. Three subsets of dysregulated genes were acquired, including 445 QDBS-specific genes, 490 YDBS-specific genes and 1676 blood stasis common genes. Our work reveals for the first time that T cell receptor, MAPK and apoptosis pathway were identified as the hub pathways based on the pathway networks, while Nfκb1, Egfr and Casp3 were recognized as the hub genes by co-expression networks. This research helps contribute to a clearer understanding of the pathological characteristics of ischemic stroke with QDBS and YDBS syndrome, the proposed biomarkers might provide insight into the accurate diagnose and proper treatment for ischemic stroke with blood stasis syndrome.

Discovery of Indazoles as Potent, Orally Active Dual Neurokinin 1 Receptor Antagonists and Serotonin Transporter Inhibitors for the Treatment of Depression.

Combination studies of neurokinin 1 (NK1) receptor antagonists and serotonin-selective reuptake inhibitors (SSRIs) have shown promise in preclinical models of depression. Such a combination may offer important advantages over the current standard of care. Herein we describe the discovery and optimization of an indazole-based chemotype to provide a series of potent dual NK1 receptor antagonists/serotonin transporter (SERT) inhibitors to overcome issues of ion channel blockade. This effort culminated in the identification of compound 9, an analogue that demonstrated favorable oral bioavailability, excellent brain uptake, and robust in vivo efficacy in a validated depression model. Over the course of this work, a novel heterocycle-directed asymmetric hydrogenation was developed to facilitate installation of the key stereogenic center.

The Novel, Nicotinic Alpha7 Receptor Partial Agonist, BMS-933043, Improves Cognition and Sensory Processing in Preclinical Models of Schizophrenia.

The development of alpha7 nicotinic acetylcholine receptor agonists is considered a promising approach for the treatment of cognitive symptoms in schizophrenia patients. In the present studies we characterized the novel agent, (2R)-N-(6-(1H-imidazol-1-yl)-4-pyrimidinyl)-4'H-spiro[4-azabicyclo[2.2.2]octane-2,5'-[1,3]oxazol]-2'-amine (BMS-933043), in vitro and in rodent models of schizophrenia-like deficits in cognition and sensory processing. BMS-933043 showed potent binding affinity to native rat (Ki = 3.3 nM) and recombinant human alpha7 nicotinic acetylcholine receptors (Ki = 8.1 nM) and agonist activity in a calcium fluorescence assay (EC50 = 23.4 nM) and whole cell voltage clamp electrophysiology (EC50 = 0.14 micromolar (rat) and 0.29 micromolar (human)). BMS-933043 exhibited a partial agonist profile relative to acetylcholine; the relative efficacy for net charge crossing the cell membrane was 67% and 78% at rat and human alpha7 nicotinic acetylcholine receptors respectively. BMS-933043 showed no agonist or antagonist activity at other nicotinic acetylcholine receptor subtypes and was at least 300 fold weaker at binding to and antagonizing human 5-HT3A receptors (Ki = 2,451 nM; IC50 = 8,066 nM). BMS-933043 treatment i) improved 24 hour novel object recognition memory in mice (0.1-10 mg/kg, sc), ii) reversed MK-801-induced deficits in Y maze performance in mice (1-10 mg/kg, sc) and set shift performance in rats (1-10 mg/kg, po) and iii) reduced the number of trials required to complete the extradimensional shift discrimination in neonatal PCP treated rats performing the intra-dimensional/extradimensional set shifting task (0.1-3 mg/kg, po). BMS-933043 also improved auditory gating (0.56-3 mg/kg, sc) and mismatch negativity (0.03-3 mg/kg, sc) in rats treated with S(+)ketamine or neonatal phencyclidine respectively. Given this favorable preclinical profile BMS-933043 was selected for further development to support clinical evaluation in humans.

40 Hz Auditory Steady-State Response Is a Pharmacodynamic Biomarker for Cortical NMDA Receptors.

Schizophrenia patients exhibit dysfunctional gamma oscillations in response to simple auditory stimuli or more complex cognitive tasks, a phenomenon explained by reduced NMDA transmission within inhibitory/excitatory cortical networks. Indeed, a simple steady-state auditory click stimulation paradigm at gamma frequency (~40 Hz) has been reproducibly shown to reduce entrainment as measured by electroencephalography (EEG) in patients. However, some investigators have reported increased phase locking factor (PLF) and power in response to 40 Hz auditory stimulus in patients. Interestingly, preclinical literature also reflects this contradiction. We investigated whether a graded deficiency in NMDA transmission can account for such disparate findings by administering subanesthetic ketamine (1-30 mg/kg, i.v.) or vehicle to conscious rats (n=12) and testing their EEG entrainment to 40 Hz click stimuli at various time points (~7-62 min after treatment). In separate cohorts, we examined in vivo NMDA channel occupancy and tissue exposure to contextualize ketamine effects. We report a robust inverse relationship between PLF and NMDA occupancy 7 min after dosing. Moreover, ketamine could produce inhibition or disinhibition of the 40 Hz response in a temporally dynamic manner. These results provide for the first time empirical data to understand how cortical NMDA transmission deficit may lead to opposite modulation of the auditory steady-state response (ASSR). Importantly, our findings posit that 40 Hz ASSR is a pharmacodynamic biomarker for cortical NMDA function that is also robustly translatable. Besides schizophrenia, such a functional biomarker may be of value to neuropsychiatric disorders like bipolar and autism spectrum where 40 Hz ASSR deficits have been documented.

Insulinotropic effect of Chikusetsu saponin IVa in diabetic rats and pancreatic ß-cells.

ETHNOPHARMACOLOGICAL RELEVANCE: As a well-known traditional Chinese medicine the root bark of Aralia taibaiensis has traditionally been used as the medicine considered alleviating several disorders including diabetes mellitus (DM). Chikusetsu saponin IVa (CHS) has been defined as a major active ingredient of triterpenoid saponins extracted from Aralia taibaiensis. The scientific evidence of anti-diabetic effect for CHS remains unknown and the purpose of our study was to study its hypoglycemic and insulin secretagogue activities. MATERIALS AND METHODS: In vivo studies were performed on type 2 diabetic mellitus (T2DM) rats given CHS for 28 days to test the antihyperglycemic activity. The in vitro effects and possible mechanisms of CHS on the insulin secretion in pancreatic ß-cell line ßTC3 were determined. RESULTS: Oral administration of CHS dose-dependently increased the level of serum insulin and decreased the rise in blood glucose level in an in vivo treatment. In vitro, CHS potently stimulated the release of insulin from ßTC3 cells at both basal and stimulatory glucose concentrations, the effect which was changed by the removal of extracellular Ca(2+). Two methods showed that CHS enhanced the intracellular calcium levels in ßTC3 cells. CHS was capable of enhancing the phosphorylation of extracellular signal-regulated protein kinases C (PKC), which could be reversed by a PKC inhibitor (RO320432), and the insulin secretion induced by CHS was also inhibited by RO320432. Further study also showed that the insulinotropic effect, intracellular calcium levels and the phosphorylation of PKC were reduced by inhibiting G protein-coupled receptor 40 (GPR40) by a GPR40 inhibitor (DC126026). CONCLUSION: These observations suggest that the signaling of CHS-induced insulin secretion from ßTC3 cells via GPR40 mediated calcium and PKC pathways and thus CHS might be developed into a new potential for therapeutic agent used in T2DM patients.

Insulin-secretagogue activity of eleven plant extracts and twelve pure compounds isolated from Aralia taibaiensis.

AIMS: To investigate the insulinogenic activities of the eleven saponins enriched traditional Chinese medicine (TCM) extracts. MAIN METHODS: Radioimmunoassay and trypan blue exclusion assay were used to investigate the insulinogenic activity and cytotoxic effects respectively. KEY FINDINGS: The total saponin extract of Aralia taibaiensis (sAT) exhibited highest insulinogenic activity and no cytotoxicity was recorded. Twelve pure compounds from sAT stimulated insulin secretion from a mouse insulinoma ßTC3 cells in a concentration-dependent manner. TA35 outperformed the other compounds which suggested that the active insulinogenic ingredient of sAT was probably TA35. In addition, both sAT and TA35 markedly potentiated glucose-induced insulin secretion. SIGNIFICANCE: Our study is the first to show that sAT dramatically stimulated insulin secretion and its antidiabetic activity may be related to its high saponin content. These findings suggested that sAT and the compound TA35 isolated from sAT may provide novel therapeutic tools for the treatment of non-insulin dependent diabetes mellitus (NIDDM).

The minimal erythema dose of broadband ultraviolet B in Taiwanese.

The purpose of this study was to determine the cutaneous response to UVB radiation in Taiwanese subjects. The relationships among minimal erythema dose (MED) of broadband UVB, Fitzpatrick's skin phototype, and skin color were examined in 61 healthy Taiwanese volunteers. Thirty-one females and 30 males were enrolled. Overall, the mean MED was 142.3 mJ/cm(2); there was no significant gender difference. The mean MEDs of subjects with Fitzpatrick skin types II, III, IV and V were 122.9 mJ/cm(2), 136.2 mJ/cm(2), 148.3 mJ/cm(2) and 165.0 mJ/cm(2), respectively. Both the mean MEDs and the lower MED limits increased with skin phototypes. To identify patients with abnormal photosensitivity, physicians could use these lower limits as reference values. In addition, different starting doses of broadband UVB phototherapy could be used for different skin phototypes.

[Curcumin attenuated the lipid peroxidation and apoptotic liver injury in copper-overloaded rats].

OBJECTIVE: Hepatolenticular degeneration (Wilson disease, WD) is an autosomal recessive disorder of copper metabolism. The clinical manifestations are dominated by the neuropsychiatric and hepatic symptoms due to copper deposition. Investigation of mechanism of copper injury should be helpful for elucidating the pathogenesis and treatment of WD. Curcumin, a plant-derived polyphenol, exhibits the properties of anti-oxidant, anti-inflammation and has no evident side effects, therefore, today curcumin is studied by more and more researchers in pharmacologic action and clinical application especially for its protective effect on liver diseases. The present study was designed to investigate the lipid peroxidation and apoptotic liver injury in copper-overloaded rats, and to explore the protective effects of curcumin. METHODS: Wistar rats, male, were randomly divided by copper-overloaded groups and curcumin treatment groups and control group. Copper-overloaded rat model was established by feeding with forage containing 1 g/kg copper sulfate and water with 0.185% copper sulfate for 8 weeks or 12 weeks. In the treatment groups, curcumin was administered orally either 50 mg/kg or 200 mg/kg for 2 weeks and 4 weeks and 8 weeks and fed with copper sulfate at the same time until the 12th week. Malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) in liver homogenates were measured to reflect the copper induced lipid peroxidation. The apoptosis of liver cell was detected by electron microscope (EM) and TUNEL assay. The expressions of TNF-alpha mRNA and IL-8 mRNA were observed by RT-PCR. Contents of TNF-alpha and IL-8 in liver homogenates were measured by ELISA. RESULTS: The MDA concentrations were significantly increased and the GSH and SOD levels were decreased in the copper-overloaded rats. The apoptosis index displayed from (2.2 +/- 1.2)% in control rats to (16.7 +/- 2.5)% in the copper treated animals. Expression of TNF-alpha mRNA and IL-8 mRNA were enhanced in the copper-overloaded rats. Curcumin significantly attenuated the increase of MDA concentrations and recovered the GSH and SOD levels. The apoptosis index decreased to (10.4 +/- 1.2)% in the copper-overloaded rats with curcumin treatment. Curcumin down-regulated the expressions of TNF-alpha mRNA and IL-8 mRNA and content of TNF-alpha and IL-8. Histological changes induced by copper in liver, such as mitochondrial swelling and endoplasmic reticulum distention and increased lysosomal granules in the model rats, were also improved significantly by curcumin treatment as evidenced by EM examination. CONCLUSION: Copper-overloading caused lipid peroxidatic injury and induced significant apoptosis in liver. TNF-alpha and IL-8 might be involved in liver injury in this model. Curcumin exhibited protective effects and possibly acted through its antioxidant and anti-apoptotic properties.