大鼠骨髓间充质干细胞分化过程的比较蛋白质组学研究

从蛋白质组学角度分析大鼠骨髓间充质干细胞(MSCs)体外定向分化为心肌细胞过程中蛋白表达情况, 采用二维电泳分离蛋白, 用PDQuest软件分析蛋白表达差异, 并采用质谱(MALDI-TOF-MS)进行鉴定, 得到了54个蛋白点, 对蛋白的生物功能分析表明, 部分蛋白通过不同的信号途径参与了MSCs的分化过程.     

Exploring peptide‐functionalized alginate scaffolds for engineering cardiac tissue from human embryonic stem cell‐derived cardiomyocytes in serum‐free medium

Engineering human cardiac tissue is a promising solution for myocardial repair of injured hearts and for drug screening. Herein, we examined the capability of chemically defined alginate scaffolds to promote cardiac tissue regeneration from human embryonic stem cell‐derived cardiomyocytes (hESC‐CMs) in serum‐free, chemically defined medium. The cells were single seeded or coseeded with human dermal fibroblasts (HFs) in macroporous scaffolds made from pristine alginate or alginate modified with arginine‐glycine‐aspartate (RGD) peptide and heparin‐binding peptide (HBP). Our results show that the addition of fibroblasts to the 3‐D culture is indispensable for the formation of functional cardiac tissues and that the presence of RGD/HBP attached to the alginate matrix further improves its functionality. The engineered tissue displayed the typical fiber morphology with massive striation. An increase in contraction amplitude and calcium transients with time, together with a decrease in excitation threshold, indicated advancement toward tissue maturation. Our results thus point to the importance of co‐cultivating fibroblasts with hESCs‐CMs in chemically defined peptide‐functionalized alginate scaffolds and culture medium for regenerating functional cardiac tissue in vitro.     

3D and Porous RGDC‐Functionalized Polyester‐Based Scaffolds as a Niche to Induce Osteogenic Differentiation of Human Bone Marrow Stem Cells

Polyester‐based scaffolds covalently functionalized with arginine‐glycine‐aspartic acid‐cysteine (RGDC) peptide sequences support the proliferation and osteogenic differentiation of stem cells. The aim is to create an optimized 3D niche to sustain human bone marrow stem cell (hBMSC) viability and osteogenic commitment, without reliance on differentiation media. Scaffolds consisting of poly(lactide‐co‐trimethylene carbonate), poly(LA‐co‐TMC), and functionalized poly(lactide) copolymers with pendant thiol groups are prepared by salt‐leaching technique. The availability of functional groups on scaffold surfaces allows for an easy and straightforward method to covalently attach RGDC peptide motifs without affecting the polymerization degree. The strategy enables the chemical binding of bioactive motifs on the surfaces of 3D scaffolds and avoids conventional methods that require harsh conditions. Gene and protein levels and mineral deposition indicate the osteogenic commitment of hBMSC cultured on the RGDC functionalized surfaces. The osteogenic commitment of hBMSC is enhanced on functionalized surfaces compared with nonfunctionalized surfaces and without supplementing media with osteogenic factors. Poly(LA‐co‐TMC) scaffolds have potential as scaffolds for osteoblast culture and bone grafts. Furthermore, these results contribute to the development of biomimetic materials and allow a deeper comprehension of the importance of RGD peptides on stem cell transition toward osteoblastic lineage.     

丝素改性聚乳酸-羟基乙酸共聚物多孔微球作为牙龈间充质干细胞递送载体的研究

利用复乳-溶剂挥发法合成适合细胞三维培养的聚乳酸-羟基乙酸共聚物(PLGA)多孔微球, 并对其表面进行丝素改性, 利用扫描电子显微镜、 能谱、 红外光谱和X射线衍射等对改性前后PLGA多孔微球的理化特性进行表征. 原代培养人牙龈间充质干细胞并进行成骨(茜素红染色)成脂(油红O染色)分化鉴定. 通过负压混悬法将牙龈干细胞负载于丝素改性的PLGA多孔微球上进行5-乙炔基-2'-脱氧尿嘧啶核苷(EdU)细胞增殖及成骨分化研究. 结果表明, 原代培养的牙龈干细胞具有多向分化潜能, 负载在丝素改性的PLGA多孔微球上的细胞有利于细胞增殖. 丝素改性的PLGA多孔微球是良好的细胞递送载体, 为进一步修复牙槽骨缺损提供了科学依据.     

大叶黄杨叶、茎、果挥发油成分及抗病毒作用

研究了大叶黄杨叶、茎、果挥发油的化学成分及抗病毒活性。 采用超临界二氧化碳萃取,应用气相色谱-质谱联用(GC-MS) 法鉴定挥发油化学成分,考察体外抗病毒作用。 共鉴定133个化合物,大叶黄杨叶挥发油中主要有2-乙氧丙烷(41.92%)、(E)-2-己烯-1-醇(17.8%)、 (E)-香叶醇(7.86%)、甲基环己烷(6.60%)等;大叶黄杨茎挥发油中主要有甲氧基苯基肟(33.10%)、二十八烷(14.34%)、α-甲基-α-[4-甲基-3-戊烯基]环氧乙烷甲醇(12.48%)、甲苯(11.88%)、二十一烷(7.74%) 等;大叶黄杨果挥发油中主要有苯甲醛(15.52%)、甲苯(15.03%)、甲基环己烷(14.76%)、(Z)-3-己烯-1-醇(10.98%)等。 大叶黄杨叶、茎、果的环己烷、乙醚萃取挥发油对特定病毒有显著抑制效果。 大叶黄杨叶、茎、果中挥发油萃取部位成分差异明显,有特定抗病毒活性。     

非定常激波三维双楔面反射的数值研究

采用基于MUSCL-Hancock插值的有限体积方法, 在非结构自适应网格上求解三维Euler方程, 研究了非定常激波在三维垂直双楔面上的反射现象. 研究结果表明, 由于三维效应的影响, 通过二维非定常激波反射理论预测的三维马赫反射区域范围存在着一定的局限, 在该区域范围内出现了一种非典型的三维突起结构, 即第二类三维马赫干. 另外, 对于不同的激波马赫数和楔面倾角组合, 非定常激波在三维双楔面上会形成具有四波结构的三维马赫反射或具有三波结构的三维规则反射.      

近红外光谱技术在天然草地植被管理中的应用

植被是监测草地健康和生产力的主要依据,其变化是草地生态系统退化和恢复状况的最直接体现。及时掌握草地植被的变化信息是防止草地退化、实现草地生态系统可持续发展的首要条件。近红外光谱技术以其简便、快捷等优越性广泛应用于不同领域,在草地资源管理中有巨大的应用潜力和前景。文章介绍了近红外光谱技术的原理、特点及其在草地植被物种组成、草地植物茎叶比、凋落物及牧草品质测定等方面的应用特点,综合阐述了近红外光谱技术在草地植被管理中的应用现状,并对其应用前景进行了展望。     

Akt Gene Therapy Enhances Angiogenesis in Ischemic Hind Limb

Both bone marrow-derived mesenchymal stem cells(MSCs) therapy and gene therapy have been applied to animal studies and clinical trials. The goal of this study was to compare the effects of three types of MSCs transplantation. In the first part of the study, MSCs were transduced with the pEGFP-C1 and transfected with the pEGFP-C1/Akt in a model of bilateral hind limb ischemia in rats. Following gene transduction, the production of Akt and VEGF protein was more in the culture media of GFP-Akt/MSCs group than that in those of the GFP/MSCs group. In the second part of the study, Sprague-Dawley rats(n=10/group) underwent surgery to create bilateral hind limb ischemia and were randomized into two groups consisting of a GFP/Akt-MSCs group(MSCs suspension, 1×107 MSCs/100 μL transfected pEGFP-C1/Akt) and a GFP-MSCs group(MSCs suspension, 1×107 MSCs/100 μL). These PEGFP-C1/Akt and PEGFP-C1 transfected MSCs suspensions were slowly infused into the adductor muscles of the rat’s left hind limb, while the rat’s right hind limb in the control group received an equal volume of PBS. Endpoints includes angiographic analysis, evaluation of capillary density, immunohistochemistry for von Willebrand factor (vWF), immunodetection of Akt and VEGF protein, RT-PCR of VEGF and Akt mRNA levels in vitro and in vivo. Our data indicate that the tissue perfusion can improve capillary density and the mature of vasculature in the GFP-Akt/MSCs group compared to that in the GFP-MSCs group or control group in the rat model of bilateral hind limb ischemia. Transplantation of MSCs transfected with Akt gene may become the future therapy for hind limb ischemia.     

Three-Dimensional Collagen Gel Networks for Neural Stem Cell-Based Neural Tissue Engineering

Stem and progenitor cells isolated from the embryonic rat cerebral cortex were immobilized by matrix entrapment in three-dimensional (3D) Type I collagen gels, and cultured in serum-free medium containing basic fibroblast growth factor. The cells trapped within the collagen networks actively proliferated and formed clone-like aggregates. Neurons were the first differentiated cells to appear within the aggregates, followed by generation of astrocytes and oligodendrocytes. In addition, necrotic cores were developed as the aggregate diameter increased and cell viability declined significantly after 3 weeks in culture. To overcome these problems, the cell-collagen constructs were transferred to Rotary Wall Vessel bioreactors for up to 10 weeks. In the rotary culture, the collagen gels compacted 3-4 folds and a long-term growth and differentiation of neural stem and progenitor cells was dynamically maintained. Remarkably, the cell-collagen constructs formed a complex two-layered structure that superficially emulated to a certain extent the cerebral cortex of the embryonic brain in architecture and functionality. The engineered 3D tissue-like constructs displaying characteristic properties of neuronal circuits may have potential use in tissue replacement therapy for injured brain and spinal cord.