摘要：
The Notch signaling pathway plays versatile roles during heart development. However, there is contradictory evidence that Notch pathway either facilitates or impairs cardiomyogenesis in vitro. In this study, we developed iPSCs by reprogramming of murine fibroblasts with GFP expression governed by Oct4 promoter, and identified an effective strategy to enhance cardiac differentiation through timely modulation of Notch signaling. The Notch inhibitor DAPT (N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester) alone drove the iPSCs to a neuronal fate. After mesoderm induction of embryoid bodies initiated by ascorbic acid (AA), the subsequent treatment of DAPT accelerated the generation of spontaneously beating cardiomyocytes. The timed synergy of AA and DAPT yielded an optimal efficiency of cardiac differentiation. Mechanistic studies showed that Notch pathway plays a biphasic role in cardiomyogenesis. It favors the early-stage cardiac differentiation, but exerts negative effects on the late-stage differentiation. Therefore, DAPT administration at the late stage enforced the inhibition of endogenous Notch activity, thereby enhancing cardiomyogenesis. In parallel, DAPT dramatically augmented the expression of Wnt3a, Wnt11, BMP2, and BMP4. In conclusion, our results highlight a practicable approach to generate cardiomyocytes from iPSCs based on the stage-specific biphasic roles of Notch signaling in cardiomyogenesis.
作者：
Liu Bai-yan;Song Xiao-ling;Yi Jian;Chen Xue-mei;Yu Yue;...
期刊：
中国结合医学杂志,2014年20(10):782-786 ISSN：1672-0415
通讯作者：
Cai Guang-xian
作者机构：
[Yi Jian; Liu Hui; Chen Xue-mei; Yu Yue; Cai Guang-xian; Liu Bai-yan; Song Xiao-ling] Hunan Univ Tradit Chinese Med, Key Lab Internal Med, Changsha 410007, Hunan, Peoples R China.
通讯机构：
[Cai Guang-xian] H;Hunan Univ Tradit Chinese Med, Key Lab Internal Med, Changsha 410007, Hunan, Peoples R China.
关键词：
Buyang Huanwu Decoction;focal cerebral ischemic;estrogen;estrogen receptor;Chinese medicine
摘要：
OBJECTIVE: To investigate the effect of Buyang Huanwu Decoction (, BYHWD) on estradiol (E2) and estradiol receptor (ER) in serum and brain in ovariectomized rats after middle cerebral artery occlusion (MCAO). METHODS: Adult female rats were ovariectomized and focal cerebral ischemic was induced by MCAO. Rats were randomly divided into normal, ovariectomy (OVX), MCAO, OVX+MCAO, OVX+MCAO+E2, and OVX+MCAO+BYHWD group. Rats were administered BYHWD 5 g/kg daily, estradiol valerate 500 mug/kg per day or distilled water for 7 consecutive days. Neuronal function and infarct volume were measured on day 7 after artery occlusion, and E2 and ER concentration in serum and brain were checked by enzyme-linked immunosorbent assay. RESULTS: BYHWD significantly improved the neurological behavior, reduced the infarction volume, increased E2 concentration in serum and brain, and increased ER concentration in the brain in ovariectomized rats after MCAO. CONCLUSION: The neuroprotective effects of BYHWD are associated with estrogen and its receptor.
期刊：
Journal of Acupuncture and Meridian Studies,2013年6(2):82-88 ISSN：2005-2901
通讯作者：
Jie Yan
作者机构：
[He, Jun-Feng; Qu, Ya-Ting] College of Traditional Chinese Medicine, Hunan University of Chinese Medicine, Changsha, China;[Yan, Jie; Li, Jiang-Shan; Wang, Chao; Chang, Xiao-Rong] College of Acu-Moxibustion and Tuina, Hunan University of Chinese Medicine, Changsha, China;[Liu, Jian-Hua] Key Laboratory of Rule and Mechanism on Acupuncture, Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou, China
通讯机构：
[Jie Yan] C;College of Acu-Moxibustion and Tuina, Hunan University of Chinese Medicine, Changsha, China
摘要：
Background: In recent years, the brain gut axis theory has received increasing attention in studies of depression. However, most studies separately address potential antidepressant and prokinetic treatments. Investigations of drugs that could potentially treat comorbid depression and gastrointestinal (GI) dysfunction via a common mechanism of action have not yet been performed in detail. Aim: To find a common mechanism of action of our patented drug, meranzin hydrate (MH), in the antidepressant and prokinetic treatment. Methods: The forced swimming test (FST) model of depression, plasma ghrelin measurement, and in vivo and in vitro measurements of GI motility were used. Results: 1. Administration of MH (9 mg/kg) decreased the immobility time during the FST after acute treatment; this effect was inhibited by the alpha 2-adrenoceptor antagonist, yohimbine, but not by the alpha 1-adrenoceptor antagonist, prazosin. 2. After chronic treatment, the immobility time of rats during the FST was decreased significantly by MH (2.25 mg/kg). 3. MH (9 mg/kg) increased plasma ghrelin levels in rats subjected to the FST; this increase was enhanced by the ghrelin receptor agonist, GHRP-6. 4. MH (9 mg/kg) also promoted gastric emptying and intestinal transit in rats with or without FST. 5. In vitro, MH (10 mu M) increased jejunal contractions in rats subjected to the FST; this effect was inhibited by yohimbine. Furthermore, the inhibitory effect of yohimbine was partly reversed by the ghrelin receptor agonist, GHRP-6. Conclusion: Our study revealed that MH from natural resources exhibits antidepressive and prokinetic-like effects through the regulation of the common mediator, the alpha 2-adrenoceptor. (C) 2012 Elsevier Ltd. All rights reserved.
摘要：
The objective of this study was to apply the "on/off" switch consisting of 3' phosphorothioate-modified allele specific primers and exo(+) polymerase in single base discrimination of A1555G and C1494T mutations in the highly conserved sites of the mitochondrial 12S rRNA. The two point mutations are the hotspot mutations associated with either aminoglycoside antibiotics induced deafness or inherited nonsyndromic hearing loss. The PCR products of mitochondrial DNA (mtDNA) 12S rRNA gene were inserted into the pMD19-T vector for transformation into Escherichia coli JM109 competent cells for preparing wild-type pMD19-T/mt vector. Inverse PCR was carried out for mtDNA 12S rRNA gene C1494T and A1555G mutagenesis and DpnI endonuclease degradating methylated pMD19-T/mt vector existing in the inverse PCR products was carried out to construct the mutation-type pMD19-T/mtM vector. These constructed vectors were confirmed by DNA sequencing. Allelic specific primers targeting wild-type and mutation-type templates were designed with 3' terminal phosphorothioate modification. Two-directional primer extension was performed using Pfu polymerases. Amplified by exo(+) polymerase, allelic specific primers perfectly matching wild-type allele were extended while no products were produced from primers targeting point-mutated deafness-related allele. Similarly, allelic specific primers perfectly matching point-mutated deafness-related mutation-type allele were extended and no products were yielded from primers targeting wild-type allele. No specific product was observed in the primer extension reaction mediated by on/off switch in screening the mtDNA 12S rRNA gene harboring either C1494T or A1555G mutation in 40 healthy volunteers tested. These data suggest that the "off switch" mediated by exo(+) polymerase is highly reliable in the diagnosis of monogenic diseases and the novel "on/off" switch has enormous applications in systematic and extended screening of the12S rRNA gene A1555G and C1494T mutations. The established assay can be widely used not only for hearing loss patients but also for normal subjects before the use of aminoglycoside antibiotics.
摘要：
Acetyl-CoA carboxylases (ACCs) play a rate-limiting role in fatty acid biosynthesis in plants, microbes, mammals and humans. ACCs have the activity of both biotin carboxylase (BC) and carboxyltransferase (CT), catalyzing carboxylation of Acetyl-CoA to malonyl-CoA. In the past years, ACCs have been used as targets for herbicides in agriculture and for drug discovery and development of human diseases, such as microbial infections, diabetes, obesity and cancer. A great number of small molecule ACC inhibitors have been developed, including natural and non-natural (artificial) products. These chemicals target BC reaction, CT reaction or ACC phosphorylation. This article provides a comprehensive review and updates of ACC inhibitors, with a focus on their therapeutic application in metabolic syndromes and malignant diseases. The patent status of common ACC inhibitors is discussed.
期刊：
Journal of Acupuncture and Meridian Studies,2012年5(2):72-79 ISSN：2005-2901
通讯作者：
Xiao-Rong Chang
作者机构：
[Yue, Zeng-Hui; He, Xin-Qun; Chang, Xiao-Rong; Liu, Mi; Fu, Ling; Zhang, Liang] College of Acu-Moxibustion and Tuina, Hunan University of Chinese Medicine, Changsha, Hunan, China;[Yuan, Jian-Ling; Yu, Bao-Sheng; Shang, Li-Chao] College of Traditional Chinese Medicine, Hunan University of Chinese Medicine, Changsha, Hunan, China
通讯机构：
[Xiao-Rong Chang] C;College of Acu-Moxibustion and Tuina, Hunan University of Chinese Medicine, Changsha, Hunan, China
作者机构：
[Zeng, Heng; Chen, Jian-Xiong] Univ Mississippi, Med Ctr, Dept Pharmacol & Toxicol, Jackson, MS 39216 USA.;[Tuo, Qinhui] Univ S China, Dept Pharmacol, Hengyang 421001, Peoples R China.;[Liao, Duan-Fang] Hunan Univ Chinese Med, Dept Tradit Chinese Diagnost, Sch Pharm, Changsha 410208, Hunan, Peoples R China.;[Chen, Jian-Xiong] Univ Mississippi, Med Ctr, Dept Pharmacol & Toxicol, 2500 N State St, Jackson, MS 39216 USA.
通讯机构：
[Chen, Jian-Xiong] U;Univ Mississippi, Med Ctr, Dept Pharmacol & Toxicol, 2500 N State St, Jackson, MS 39216 USA.
摘要：
Diabetes is associated with impairment of angiogenesis such as reduction of myocardial capillary formation. Our previous studies demonstrate that disruption of Angiopoietin-1 (Ang-1)/Tie-2 signaling pathway contributes to the diabetes-associated impairment of angiogenesis. Protein tyrosine phosphatase (PTP) has a critical role in the regulation of insulin signal by inhibition of tyrosine kinase phosphorylation. In present study, we examined the role of protein tyrosine phosphatase-1 (SHP-1) in diabetes-associated impairment of Ang-1/Tie-2 angiogenic signaling and angiogenesis. SHP-1 expression was significantly increased in diabetic db/db mouse hearts. Furthermore, SHP-1 bond to Tie-2 receptor and stimulation with Ang-1 led to SHP-1 dissociation from Tie-2 in mouse heart microvascular endothelial cell (MHMEC). Exposure of MHMEC to high glucose (HG, 30 mmol/L) increased SHP-1/Tie-2 association accompanied by a significant reduction of Tie-2 phosphorylation. Exposure of MHMEC to HG also blunted Ang-1-mediated SHP-1/Tie-2 dissociation. Knockdown of SHP-1 significantly attenuated HG-induced caspase-3 activation and apoptosis in MHMEC. Treatment with PTP inhibitors restored Ang-1-induced Akt/eNOS phosphorylation and angiogenesis. Our data implicate a critical role of SHP-1 in diabetes-associated vascular complications, and that upregulation of Ang-1/Tie-2 signaling by targeting SHP-1 should be considered as a new therapeutic strategy for the treatment of diabetes-associated impairment of angiogenesis.
