Rosmarinic acid elicits neuroprotection in ischemic stroke via Nrf2 and heme oxygenase 1 signaling.

Rosmarinic acid (RA) can elicit a neuroprotective effect against ischemic stroke, but the precise molecular mechanism remains poorly understood. In this study, an experimental ischemic stroke model was established in CD-1 mice (Beijing Vital River Laboratory Animal Technology, Beijing, China) by occluding the right middle cerebral artery for 1 hour and allowing reperfusion for 24 hours. After intraperitoneally injecting model mice with 10, 20, or 40 mg/kg RA, functional neurological deficits were evaluated using modified Longa scores. Subsequently, cerebral infarct volume was measured using TTC staining and ischemic brain tissue was examined for cell apoptosis with TUNEL staining. Superoxide dismutase activity and malondialdehyde levels were measured by spectrophometry. Expression of heme oxygenase-1 (HO-1), nuclear factor erythroid 2-related factor 2 (Nrf2), Bcl-2, Bax, Akt, and phospho-Ser473 Akt proteins in ischemic brain tissue was detected by western blot, while mRNA levels of Nrf2, HO-1, Bcl-2, and Bax were analyzed using real time quantitative PCR. In addition, HO-1 enzyme activity was measured spectrophotometrically. RA (20 and 40 mg/kg) greatly improved neurological function, reduced infarct volume, decreased cell apoptosis, upregulated Bcl-2 protein and mRNA expression, downregulated Bax protein and mRNA expression, increased HO-1 and Nrf2 protein and mRNA expression, increased superoxide dismutase activity, and decreased malondialdehyde levels in ischemic brain tissue of model mice. However, intraperitoneal injection of a HO-1 inhibitor (10 mg/kg zinc protoporphyrin IX) reversed the neuroprotective effects of RA on HO-1 enzyme activity and Bcl-2 and Bax protein expression. The PI3K/Akt signaling pathway inhibitor LY294002 (10 mM) inhibited Akt phosphorylation, as well as Nrf2 and HO-1 expression. Our findings suggest that RA has anti-oxidative and anti-apoptotic properties that protect against ischemic stroke by a mechanism involving upregulation of Nrf2 and HO-1 expression via the PI3K/Akt signaling pathway.

Apelin-13 protects against apoptosis by activating AMP-activated protein kinase pathway in ischemia stroke.

Apelin has been proved to be protective against apoptosis induced by ischemic reperfusion. However, mechanisms whereby apelin produces neuroprotection remain to be elucidated. AMP-activated protein kinase (AMPK) is a master energy sensor that monitors levels of key energy metabolites. It is activated via AMPKαThr172 phosphorylation during cerebral ischemia and appears to be neuroprotective. In this study, we investigated the effect of apelin on AMPKα and tested whether apelin protecting against apoptosis was associated with AMPK signals. Focal transient cerebral ischemia/reperfusion (I/R) model in male ICR mice was induced by 60 min of ischemia followed by reperfusion. Apelin-13 was injected intracerebroventricularly 15 min before reperfusion. AMPK inhibitor, compound C, was injected to mice intraperitoneally at the onset of ischemia. In experiment 1, the effect of apelin-13 on AMPKα was measured. In experiment 2, the relevance of AMPKα and apelin-13' effect on apoptosis was measured. Data showed that apelin-13 significantly increased AMPKα phosphorylation level after cerebral I/R. Apelin-13, with the co-administration of saline, reduced apoptosis cells, down-regulated Bax and cleaved-caspase3 and up-regulated Bcl2. However, with the co-administration of compound C, apelin-13 was inefficient in affecting apoptosis and Bax, Bcl2 and cleaved-caspase3. The study provided the evidence that apelin-13 up-regulated AMPKα phosphorylation level in cerebral ischemia insults and AMPK signals participated in the mechanism of apelin-mediated neuroprotection.

Development of highly sensitive amperometric biosensor for glucose using carbon nanosphere/sodium alginate composite matrix for enzyme immobilization.

In this study, we developed a highly sensitive amperometric biosensor for glucose detection based on glucose oxidase immobilized in a novel carbon nanosphere (CNS)/sodium alginate (SA) composite matrix. This hybrid material combined the advantages of CNS and natural biopolymer SA. This composite film was characterized by scanning electron microscope, electrochemical impedance spectroscopy and UV-vis, which indicated that the hybrid material was suitable for immobilization of glucose oxidase. Various experimental conditions were investigated that influenced the performance of the biosensor, such as pH, applied potential and temperature. Under the optimum conditions, the biosensor showed excellent performance for glucose over a wide linear concentration range from 1.0 × 10(-6) to 4.6 × 10(-3) M with a detection limit of 0.5 µM based on a signal-to-noise ratio of 3. Furthermore, the biosensor exhibited excellent long-term stability and satisfactory reproducibility.

[Comparison of Dongeuisoosebowon sasung caobenjuan (The Four Constitutional Medicine of A Draft Volume of Longevity and Life Preservation in Eastern Medicine) and Dongeuisoosebowon (Longevity and Life Preservation in Eastern Medicine)].

In 1894, Lee Je-ma finished the book Dongeuisoosebowon sasung caobenjuan (The Four Constitutional Medicine of A Draft Volume of Longevity and Life Preservation in Eastern Medicine), and later, in 1900, he started revising the book. However, he passed away and didn't finish his work. In 1901, Lee's disciples edited the revised edition, and published it in the title Dongeuisoosebowon (Longevity and Life Preservation in Eastern Medicine). Some hand-copied versions of Dongeuisoosebowon sasung caobenjuan were still available. After comparing these two books, it was found that Lee's disciples made rather more changes: including addition of its contents, adjustment of its order, modification of the texts and prescriptions, etc. By comparison, the contents in the hand-copied edition more closely reflects Lee's recognition on the Sasung Euihak (Four-Constitution medicine), while those of the published ones were deleted a bulk of the original contents of the Four-Constitutional medicine, yet supplemented much contents of traditional Chinese medicine.