人脂肪组织来源干细胞在心脏疾病治疗中的应用

目的:综述人脂肪组织来源干细胞的生物学特性及其在缺血性心脏病中的应用,分析不足,并在此基础上提出未来研究要解决的问题,以期为临床治疗提供依据。资料来源:应用计算机检索Blackwell、Elsevier、Pubmed数据库1980/2007期间脂肪源性干细胞与缺血性心脏病方面的文献,检索词为“bone mesenchymal stem cells,adipose derived stemcells,cardiomyocytes,ischemic heart disease”等。应用计算机检索中国期刊全文数据库1980/2007期间相关文献,检索词为“骨髓间充质干细胞,脂肪组织来源的干细胞,心肌细胞,缺血性心脏病”等。并手工查阅相关书籍。资料选择:对资料进行初步选择:①脂肪组织来源干细胞的生物学特性。②脂肪组织来源干细胞治疗缺血性心脏病。排除重复文献。资料提炼:共搜集到相关文章57篇,删除内容重复及与本文主题关系较远的文章,剩余41篇作为综述参考。资料综合:脂肪组织来源干细胞与同样起源于中胚层的骨髓基质细胞不仅具有非常相似的生物学特性,而且在细胞表面标志谱的表达方面也非常相近。并且脂肪组织来源广泛,取材方便,可获得的基质细胞数量大,易于培养扩增。有研究发现,脂肪组织来源干细胞体外培养不需要任何诱导便能分化成具有自律性的心肌细胞,使得脂肪组织来源干细胞治疗缺血性心脏病成为可能。结论:脂肪组织来源干细胞在取材和增殖方面较骨髓间充质干细胞有优势;脂肪组织来源干细胞能较好的诱导为心肌细胞,将为缺血性心脏病的治疗提供更广阔的前景。     

人骨髓间充质干细胞分离培养及血管内皮生长因子对其产生的诱导分化作用

目的：目前有关骨髓间充质干细胞向内皮细胞诱导分化的研究较少。本实验分离和培养人骨髓间充质干细胞，用带有VEGF165的质粒转染人骨髓间充质干细胞，探讨血管内皮生长因子对其体外诱导分化的作用。 方法：实验于2005—04／2006—04在吉林大学人兽共患病教育部重点实验室完成。取成人的已排除血液系统肿瘤疾病的新鲜骨髓（自愿提供），采用Percoll梯度分离培养骨髓间充质干细胞，于倒置显微镜下观察细胞形态变化和生长情况。原代细胞培养至增殖接近融合状态时，单克隆培养法分离传代培养，扩增骨髓间充质干细胞。采用流式细胞术检测细胞免疫学表型。在原核细胞大肠杆菌DH5α中复制扩增和提取，纯化、克隆pcDNA3．0-VEGF165质粒。用脂质体转染法转染骨髓间充质干细胞：应用流式细胞术检测诱导后骨髓间充质干细胞免疫学表型变化j并采用免疫荧光染色鉴定转染情况，并设质粒空载和未转染的骨髓间充质干细胞为对照。 结果：人骨髓间充质干细胞原代培养1周后，造血细胞消失，贴壁细胞体积增大，呈现梭形外观，有粗大的细胞突起伸出。2周后细胞融合成单层，梭形突起变长，排列有明显的方向性，细胞排列成旋涡状、网状、辐射状。流式细胞术显示，人骨髓间充质干细胞免疫学表型CD44、CD29阳性，CD34、CD31、CD45阴性。VEGF165诱导骨髓间充质千细胞后CD44表达明显降低，CD31明显升高。免疫荧光染色显示，用FITC标记后的VEGF抗体使细胞显现绿色荧光，用cy3标记的CD31抗体使细胞显现了红色荧光。 结论：转染后的骨髓间充质干细胞细胞表型发生明显转变，CD31表达率明显增高，呈现典型的内皮细胞的表型特征，这说明骨髓间充质干细胞具有向内皮细胞分化的潜能。     

Altering dietary lysine:arginine ratio has little effect on cardiovascular risk factors and vascular reactivity in moderately hypercholesterolemic adults

BackgroundInformation is scarce regarding the effect of dietary protein type, with specific focus on the lysine-to-arginine (Lys:Arg) ratio, on cardiovascular risk factors and vascular reactivity in humans.ObjectiveDetermine the effect of dietary Lys:Arg ratio on cardiovascular risk factors and vascular reactivity in moderately hypercholesterolemic adults.DesignRandomized cross-over design of two 35-day diet phases; thirty adults (21 females and 9 males, ≥50 years, LDL cholesterol ≥120 mg/dL). Diets had 20% energy (E) protein, 30% E fat, 50% E carbohydrate and were designed to have low (0.7) or high (1.4) Lys:Arg ratio. Measures included fasting and postprandial lipid, lipoprotein, apolipoprotein concentrations; fasting high sensitivity C-reactive protein (hsCRP), small dense LDL (sdLDL) cholesterol, remnant lipoprotein cholesterol (RemLC), glycated albumin, adiponectin and immunoreactive insulin concentrations, endogenous cholesteryl ester transfer protein (CETP) and lecithin:cholesterol acyl transferase (LCAT) activities; cholesterol fractional synthesis rate (FSR); and flow mediated dilation (FMD) and peripheral artery tonometry (PAT).ResultsNo differences were observed in fasting and/or postprandial total, LDL, HDL and sdLDL cholesterol, RemLC, Lp(a) or apo B concentrations, LCAT and CETP activities, FSR, glycated albumin, immunoreactive insulin, FMD or PAT. The low, relative to the high, Lys:Arg ratio diet resulted in lower postprandial VLDL cholesterol (?24%, P = 0.001) and triglycerides (?23%, P = 0.001), and small but significant differences in fasting (?3%, P = 0.003) and postprandial (?3%, P = 0.018) apo AI, and fasting adiponectin concentrations (+7%, P = 0.035). Fasting and postprandial hsCRP concentrations were 23% lower after the low Lys:Arg ratio diet (P = 0.020 for both).ConclusionsDiets differing in Lys:Arg ratios had no or small effects on cardiovascular risk factors and vascular reactivity.     

关于全基因组相关分析的研究--你首先想要什么：好消息,坏消息,或者好消息？

很久以来都认为遗传成分参与2型糖尿病（T2DM）的发病，但是有关遗传学上的发现一直进展缓慢，这是由于遗传成分的复杂性。有关糖尿病相关的各种表型的研究的大量资料提示，所谓的“T2DM”可能是许多疾病的统称，由于它们具有通常相互重叠的多种基本发病机制。因此，对曾抱有期望的T2DM的遗传学基础的寻求已经证明是很艰难的。     

Purified murine hematopoietic stem cells function longer on nonirradiated W41/Wv than on +/+ irradiated stroma

Mouse bone marrow (BM) was separated into low-density, lineage- negative, wheat germ agglutinin-positive (WGA+), Rhodamine-123 bright (Rhbright) or dim (Rhdim) cells to obtain populations that were highly enriched for committed progenitors (Rhbright cells) or for more primitive stem cells (Rhdim). When 2,500 Rhbright or Rhdim cells were seeded onto 6-week-old irradiated (20 Gy) long-term BM cultures (LTBMC), the nonadherent cell production from Rhbright cells was transient and ended after 5 weeks. Production from Rhdim cells did not begin until week 3, peaked at week 5, and ended at week 8, when the irradiated stroma seemed to fail. Termination of cell production from Rhdim cells did not occur in nonirradiated LTBMC from W41/Wv mice. During peak nonadherent cell production, 25% to 30% of the cells in the nonirradiated LTBMC from W41/Wv mice had donor cell markers. Two approaches were tested to try to enhance the proportion or number of donor cells. Addition of Origen-HGF at the time of seeding Rhdim cells caused a nonspecific increase in both host and donor cell production, but a specific increase in production of donor cells was obtained by seeding the cultures at 2 weeks rather than 6 weeks. Limiting dilution of Rhdim cells gave the same frequency of wells producing cells on both irradiated +/+ and nonirradiated W41/Wv or W/Wv cultures.     

Stability of mitomycin admixtures

The stability of mitomycin in admixtures for continuous intravenous infusion was studied. Mitomycin was reconstituted and diluted to 50 micrograms/mL in polyvinyl chloride minibags containing 5% dextrose injection 50 mL or 0.9% sodium chloride injection 50 mL. Additional mitomycin admixtures were reconstituted with a buffer solution containing monobasic and dibasic sodium phosphate; these were diluted with 5% dextrose injection only. Admixtures were stored at room temperature (27-30 degrees C) and refrigerated temperature (5 degrees C) for 120 days. Mitomycin concentrations in each admixture were tested by high-performance liquid chromatography (HPLC) immediately after admixture and at intervals during storage. Ultraviolet spectra were determined at the same time as HPLC analysis, and the admixtures were visually inspected and tested for pH. Mitomycin concentrations decreased rapidly in the unbuffered admixtures; after 12 hours at room temperature, less than 26% of the drug remained in the dextrose admixture. When the unbuffered admixtures were refrigerated for 12 hours, the mitomycin concentrations decreased 10% in the sodium chloride admixtures and 33% in the dextrose admixtures; after 24 hours, the percentages of drug loss were 23% and 42%, respectively. Mitomycin concentrations in the buffered admixtures showed no substantial decrease during 120 days at 5 degrees C. At room temperature, concentrations decreased 10% after 15 days. When the admixture is buffered to a pH of approximately 7.8, mitomycin is stable in 5% dextrose injection for up to 15 days at room temperature and at least 120 days at 5 degrees C. Unbuffered mitomycin admixtures should not be stored or administered by prolonged i.v. infusion.     

A new combined in-vitro test model for the identification of substances affecting essential sperm functions [published erratum appears in Hum Reprod 1997 Nov;12(11):2580]

Binding of mammalian spermatozoa to the zona pellucida and the induction of the acrosome reaction are prerequisites for successful oocyte fertilization. It has been postulated that xenobiotics that are released in the environment as well as exposure to pharmaceutical medications may be associated with reproductive problems in men and wildlife. Examining physiological and non-physiological effects of particular compounds on sperm functions requires high quality in-vitro test systems. We established a reliable combined in-vitro test system with bovine gametes and evaluated if aliquots of pooled post-thaw spermatozoa are suitable for examining essential sperm functions. Using cryopreserved semen, the PSA-FITC/Hoechst 33258 staining procedure was applicable to evaluate the acrosomal status and cell viability. In the bovine hemizona assay, hemizona indices revealed no differences between cryopreserved and fresh semen. Treatment of post-thaw bovine spermatozoa with progesterone (1 microM or bovine follicular fluid (20%) induced the acrosome reaction from 12% (untreated spermatozoa) to 25% (P < 0.001) and to 22% [corrected] (P < 0.01), respectively. Incubation of both compounds (1 microM progesterone and 20% follicular fluid) raised the percentage of acrosome-reacted spermatozoa to 30% (P < 0001). Our results demonstrate that cryopreserved semen can be integrated into an in-vitro screening model for reproductive toxicology testing. Pooled, cryopreserved bovine spermatozoa will thus permit reproducible experiments for clinical and basic science purposes and may also be applicable for the human system.