RegeneRAtive medicine for bRAin and nerve repair
We isolated and propagated neuRAl stem cells from the exposed bRAin tissue of the patients with open bRAin tRAuma, and then implanted neuRAl stem cells with MRI-guided stereotactic device for the patients. Within 2-years follow-ups, the patients were investigated for functional recovery. ContRAst to the case control group, implantation of neuRAl stem cells was associated with a significant improvement in patient's neurological function. Investigations of stem cell theRApy have required analysis of the fate and migRAtion of implanted neuRAl stem cells. Here, We demonstRAte the feasibility of labeling human neuRAl stem cells and retinal stem cells with nanoparticle and tRAcking of implanted cells in monkey and human centRAl nervous system (CNS). This data demonstRAtes the possibility of stem cell theRApy in CNS and collectively provide necessary foundation for overcoming challenges to the enhancement of tRAnslational regeneRAtive medicine of bRAin and optic nerve injury.
sRNA Induces the Large-scale TRAnsdetermination of Mesenchymal Stem Cells into Hematopoietic Stem Cells in Human.
Mesenchymal stem cells (MSCs) can differentiate into cells of bone, endothelium, adipose tissue, cartilage, muscle, and bRAin. However, whether they can tRAnsdeterminate into hematopoietic stem cells (HSCs) remains unsolved. We report here that a subpopulation of human MSCs that are CD44+,CD29+, CD105+, CD166+,CD133-,CD34- could differentiate into hematopoietic stem cells (CD150+/CD133+/CD34+) and their descending blood cells in vitro, when tRAnsfected with new endogenous shRNAs The sRNA was high-effectively delivered into MSCs by a novel peptide means. These induced MSC-HSCs could form different types of hematopoietic colonies as nature-occurring HSCs did. Upon tRAnsplantation into sublethally irRAdiated NOD/SCID mice, these MSC-HSCs engRAfted and differentiated into all hematopoietic lineages such as erythrocytes, lymphocytes, myelocytes and thrombocyte. More importantly, these induced HSCs could successfully engRAft and effectively function in patients with severe aplastic anemia. Furthermore, we demonstRAted the first evidence that the tRAnsdetermination of MSCs was induced by acetylation of histone proteins and activation of many tRAnscriptional factors. Together, our findings identify the sRNAs that dictates a directed differentiation of MSCs toward HSCs and open up a new source for HSCs used for the treatment of blood diseases and artificial stem cell-made blood.
GE公司His标签蛋白纯化预装柱HisTRAp™ FF Crude使用技巧
主要介绍了His标签蛋白纯化预装柱HisTRAp™ FF Crude的实验过程，原理，说明，一般事项，样品制备，纯化操作， 按比例放大， 柱的清洗及保存等。
immunotheRApy against cancers(position of sellular theRApy)
RegeneRAtive medicine for bRAin and nerve repair
We isolated and propagated neuRAl stem cells from the exposed bRAin tissue of the patients with open bRAin tRAuma, and then implanted neuRAl stem cells with MRI-guided stereotactic device for the patients. Within 2-years follow-ups, the patients were investigated for functional recovery. ContRAst to the case control group, implantation of neuRAl stem cells was associated with a significant improvement in patient's neurological function. Investigations of stem cell theRApy have required analysis of the fate and migRAtion of implanted neuRAl stem cells. Here, We demonstRAte the feasibility of labeling human neuRAl stem cells and retinal stem cells with nanoparticle and tRAcking of implanted cells in monkey and human centRAl nervous system (CNS). This data demonstRAtes the possibility of stem cell theRApy in CNS and collectively provide necessary foundation for overcoming challenges to the enhancement of tRAnslational regeneRAtive medicine of bRAin and optic nerve injury.
Targets-based theRApy for leukemia: opportunity and challenge
优点主要为：能增强患者的免疫力，防止肿瘤的转移和复发，对病人机体的损伤小。 在我国，现在普遍开展的树突状细胞（DC）和细胞因子诱导的杀伤细胞（CIK）的生物疗法被广泛应用。
Erich Gnaiger：Life Style and Mitochondrial Competence – Modern Drugs for T2 Diabetes in Aging and DegeneRAtive Diseases.
D. Swarovski Research LaboRAtory (Mitochondrial Physiology), Dept. GeneRAl, VisceRAl and TRAnsplant Surgery, Innsbruck Medical University; and OROBOROS INSTRUMENTS, Innsbruck, Austria. - Email: erich.gnaiger@oroboros.at
The contribution of mitochondrial dysfunction to the etiology of T2 diabetes and a RAnge of preventable metabolic diseases is the subject of intensive current research with world-wide health implications.
Recently these investigations gained depth and scope by technological advances for diagnosis of mitochondrial function by comprehensive OXPHOS analysis using high-resolution respirometry [1,2]. Fundamental questions of a causal relationship, however, between compromised mitochondrial function and development of T2 diabetes remain to be resolved [3,4] to optimize prevention and treatment of insulin resistance.
For preventable diseases such as T2 diabetes, the evolutionary background of mitochondrial competence provides a solid basis for improved and broad application of a well established modern drug, mtLSD.
Post-industrial societies are chaRActerized by a high-energy input lifestyle with diminished physical activity and high incidence of non-tRAnsmittable diseases, in comparison to human populations where physical work is essentially important for sustaining life and in which degeneRAtive diseases (T2 diabetes, various cancers, Alzheimer's) are essentially absent [5]. The capacity of oxidative phosphorylation (OXPHOS) is increased or maintained high by a life style involving enduRAnce exercise and strength tRAining [6].
Life style changes from the age of 20-30 years to the elderly, but is subject to change and intervention. Depending on group selection in cross-sectional studies, OXPHOS capacity declines from the age of 20-30 years [7,8], or is independent of age up to 80 years [9,10].
Independent of age, there is a strong decline of OXPHOS capacity in human vastus lateRAlis from BMI of 20 to 30 [1]. At a BMI >30, a threshold OXPHOS capacity is reached in human v. lateRAlis that may be chaRActeristic of a low-gRAde inflammatory state (‘mitochondrial fever’).
Onset of degeneRAtive diseases (T2 diabetes, neuromuscular degeneRAtion, various cancers) and mitochondrial dysfunction inteRAct in an amplification loop progressing slowly with age, such that cause and effect of mitochondrial dysfunction cannot be distinguished. Diminished antioxidant capacity at low mitochondrial density is an important mechanistic candidate in the state of mitochondrial fever.
For implementing a life style supporting mitochondrial competence and preventing degeneRAtive diseases in modern societies, we need (1) extended research progRAmmes focused on the causative link between mitochondrial competence and effective prevention of degeneRAtive diseases, (2) educational progRAmmes on mitochondrial physiology targeted at geneRAl pRActitioners, teachers and the society at large, (3) coopeRAtion of health care and insuRAnce organizations to support preventive life style activities, and (4) do not miss any opportunity in taking the lead in living the mtLife Style Drug (mtLSD).
秦正红：DRAM1 regulates autophagy flux and Bid-mediated cell death via lysosomes
秦正红，博士，教授，神经药理专业博士生导师。1994年在美国宾州医学院研究生院获博士学位，先后在美国国家卫生研究院(NIH)及麻省总医院和哈佛大学医学院从事研究工作。2003年从哈佛大学引进，现为苏州大学医学部基础医学与生物科学学院科研中心实验室主任，中国药理学会生化药理学专业委员会委员，中国药理学会神经药理学专业委员会委员，美国神经科学学会会员。
Damage-regulated autophagy modulator1 (DRAM1), a novel TP53 target gene, is an evolutionarily conserved lysosomal protein and plays an essential role in TP53-dependent autophagy activation and apoptosis (Crighton et al, 2006). However, the mechanisms by which DRAM1 promotes autophagy and apoptosis are not clear.
3-Nitropropionic acid (3-NP) is an inhibitor of mitochondrial respiRAtory complex II. IntRAstriatal administRAtion of 3-NP produces neuropathology resemble to Huntington disease. 3-NP-induced neuronal death was involved in autophagy and apoptosis. In vitro studies with 3-NP in TP53 wt and null cells, 3-NP or CCCP increased the protein levels of DRAM1 in a TP53-dependent or independent manner. DRAM1 induction contributed to 3-NP-induced autophagy activation. Knock-down of DRAM1 with siRNA inhibited the activity of V-ATPase, acidification of lysosomes and activation of lysosomal proteases. Knock-down of DRAM1 reduced the cleaRAnce of autophagososmes.
3-NP also induced a tRAnscription independent upregulation of BAX protein levels. Knock-down of DRAM1 suppressed the increase in BAX levels. Co-immunoprecipitation and pull-down studies revealed an inteRAction of DRAM1 and BAX protein. Stably expression of exogenous DRAM1 increased the half-life of BAX. Upregulation of DRAM1 recruited BAX to lysosomes and induced cathepsin B-dependent cleavage of Bid and cytochrome c release. Knockdown of DRAM1, BAX or inhibition of lysosomal enzymes reduced 3-NP-induced cytochrome c release and cell death.
These data suggest that DRAM1 plays important roles in regulating autophagy flux and apoptosis. DRAM1 promotes autophagy flux through a mechanism involves activation of V-ATPase and enhances the acidification of lysosomes. DRAM1 promotes apoptosis via a mechanism involving recruitment of BAX to lysosomes to trigger cathepsin B-mediated Bid cleavage.
GeneRAting B-lymphoblastoid cell lines using Epstein Barr virus tRAnsformation.
GeneRAting immortalized B-lymphoblastoid cell lines via Epstein Barr virus tRAnsformation using the B95-8 EBV-infected and producing marmoset cell line.