Bone marrow-derived stromal cells and cartilage tissue engineering

Articular cartilage repair is one of the most challenging issues which remains to be resolved in clinic work. Discovering of bone marrow stromal cells (BMSC) and its application in tissue engineering provide new methods for the treatment of cartilage defects. High seeding density, appropriate cytokines and three-dimensional culture play important roles in the process of inducing BMSC differentiating into chondrocytes, suitable scaffold is also essential in reconstructing cartilage in vitro by methods of tissue engineering.     

Selection and amplification of the liver stem cell subset from rat bone marrow cells with a medium containing cholestatic serum in vitro

AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro. METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.     

Effects of TGF-β1 and TNF-α antisense PS-ODNS on ex vivo expansion of hematopoietic stem/progenitor cells(HSPC)

AIM:To study the effect of TGF-β₁ and TNF-α antisense PS-ODNS on ex vivo expansion of hematopoietic stem/progenitor cells (HSPC). METHODS:CD₃₄⁺cells were purified from fresh umbilical cord blood by immunomagnetic beads, and mononuclear cells were purified from bone marrow by Ficoll-hypaque. The effects of TGF-β₁ and /or TNF-α antisense PS-ODNS on ex vivo expansion of CD₃₄⁺ cells、CFU-GEMM、CFU-GM、CFU-E and BFU-E were detected by using liquid and semi-solid culture systems.RESULTS:TGF-β₁ antisense PS-ODNS cooperated with cytokines increased the number of CD₃₄⁺ cells, CFU-GEMM, CFU-GM, CFU-E and BFU-E, which was as 4, 2.6, 2.7, 1.8, 2.1 times as that of the control (the cytokines combination), respectively. TNF-α antisense PS-ODNS cooperated with cytokines respectively increased the number of CD₃₄⁺ cells, CFU-GEMM, CFU-GM, CFU-E and BFU-E by 4, 2.9, 2.6, 1.7, 1.8 times as that of the control. The above two antisense PS-ODNS cooperated with cytokines could respectively increased the number of CD₃₄⁺ cells, CFU-GEMM, CFU-GM, CFU-E and BFU-E by 5.3, 2.1, 2.7, 1.9, 1.8 times as that of the control.CONCLUSION:Inhibition of endogenous TGF-β₁ and TNF-α by antisense PS-ODNS will be one of the effective methods to expand HSPC ex vivo.     

Effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts

AIM: To investigate the effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts.METHODS: hMSC were separated from human marrow and expanded in cuture medium. hMSC were induced with dexamethasone, β-glycerophosphate, vitamin C which acted as osteoblast differentiation inducer. PD98059 was added into the osteoblasts induction medium. The cells were assayed with cell morphology, alkaline phosphatase (AP) activity and calcium deposition. RESULTS: The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. After induced with osteoblasts induction medium, the cells showed changes in cell morphology from spindle-shape to cuboidal and polygonal. The AP activity increased gradually and reached the peak in 12 days, then decreased. Many scattered tangerice calcium nodes were observed. PD 98059 significantly inhibited AP activity and calcium deposition in a dose-dependent manner. A striking observation of the present study was that a few adipocytes appeared in cultures that were treated with PD 98059 and osteogenic differentiation medium. CONCLUSION: These results indicated that osteogenic diferentiation from the hMSCs was related to the activation of the ERK.     

Effects of Taoren Honghua Jian on the hyperlipidemia without symptom

AIM: To examine the effect of traditional chinese medicine recipe, Taoren Honghua(semen persicae-flos carthami) decoction, on hyperlipidemia without symptom. METHODS: The plasma TC, TG, LDL, HDL, apolipoprotein(Apo) A, Apo Bof the patients with hyperlipidemia without symptom were measured using automatic analyzer (shimadiu CL-7200), the production of nitric oxide(NO) was detected by Greiss reaction, and SODactivity and MDAformation were examined using o-trihydroxy benzene and barbituric methotheds, respectively. RESULTS: After oral administration of Taoren Honghua decoction, the plasma levels of TC, TG, LDL, and MDAof the patients were markedly decreased, however, the plasma levels of ApoA, HDL, SODand NOwere significantly increased and almost no change was detected in the plasma level of Apo B. In control group, it was found that although the plasma level of TC, TG and LDL were decreased ( P<0.05 ) and ApoAas well as HDLwere increased, ApoB, SOD, MDA and NO production were all unchanged. CONCLUSION: The traditional chinese medicine recipe, Taoren Honghua decoction,has a significant therapeutic effect on patients with hyperlipidemia by removing blood stasis, promoting qi circulation and in turn reducing blood lipid level.     

引起糖甜菜细菌性叶斑病的萎蔫短小杆菌新致病变种

 1995年在内蒙古临河市新发现了糖甜菜细菌性叶斑病,从病斑所分离的10个细菌菌株经柯赫氏法则验证,均确系该病的病原菌。采用形态观察、表型特征和生理生化特性测定、数值分析、血清学反应、细胞化学成分分析、DNA G+C mol%和DNA-DNA同源性测定进行了鉴定,并与植物病原棒形细菌15个标准菌株进行了比较。该病原菌为革兰氏阳性细菌,不规则短杆状,有一根鞭毛、亚极生或侧生,结合其生理生化特性、细胞化学成分和DNA G+C mol%和DNA-DNA同源性测定结果,认为应属于短小杆菌属(Curtobacterium)的萎蔫短小杆菌(Cur. flaccumfaciens),数值分析也支持这一结论。此外,据血清学反应结果及其对短小杆菌属的其它植物寄主的致病情况,认为该病原菌应是萎蔫短小杆菌种下的一个新的致病变种,定名为Curtobacterium flaccumfaciens pv. beticola pv. nov. Chen et al.,2000(萎蔫短小杆菌糖甜菜致病变种)。     

粪臭素在家畜养殖中的危害

粪臭素是L-色氨酸的细菌降解产物，是引起反刍动物急性肺水肿和肺气肿(ABPE)的主要物质，同时也是产生猪肉异臭味的主要物质，严重危害家畜养殖。本文综述了粪臭素的理化性质及合成途径、在家畜中的作用机理、影响粪臭素的因素及降低家畜体内粪臭素的方法等内容。     

植酸酶在饲料中的研究与应用

植酸酶在饲料中应用，不仅能提高植酸磷的消化率，减少配方中无机磷的添加量和磷排泄污染，还可改善和提高蛋白质、能量、氨基酸和微量元素的利用率。     

猕猴桃果实成熟前补钙对果实含钙量的影响

猕猴桃果实成熟前用喷叶法和喷果法补钙，可显著提高果实中的钙含量。喷叶较喷果效果好。用Ca(NO3)2喷叶，比用CaCl2处理效果好，差异显著。而果实对CaCl2和Ca(NO3)2的吸收选择性差异不显著。经补钙处理的果实采后进行贮藏效果观察发现：用Ca(NO3)2喷叶处理的果实贮藏性较好，用其喷果处理次之；用CaCl2喷果和喷叶处理的果实贮至28天软果率均达到90％。     

利用远缘杂交创造核果类果树新种质的研究

以桃、杏、李、樱桃、杏梅等果树的 1 4个品种为亲本 ,3年来共进行了 80余个组合的远缘杂交试验 ,探讨了核果类果树远缘杂交的亲和性 ,并对远缘杂种幼胚进行了胚抢救。结果表明 ,铃铛花期授粉的坐果率显著高于初花期授粉 ;同一杂交组合 ,正反交坐果率差异显著 ,母本对远缘杂交亲和性影响很大 ,选择自交亲和或自然坐果率高的种或品种做母本容易克服远缘杂交的不亲和性 ;适宜场强的静电场、He -Ne激光处理及60 Coγ射线与He -Ne激光联合处理花粉 ,均能显著提高花粉离体萌芽率 ,用上述处理的花粉进行的远缘杂交坐果率也明显高于对照 ;60 Coγ射线单独处理则降低了花粉离体萌芽率 ,远缘杂交的坐果率也低于对照 ;对远缘杂种幼胚及时地进行胚抢救 ,并诱导形成多丛芽 ,是克服核果类果树远缘杂种不育性的有效方法。研究并筛选出了李、樱桃胚萌发与生长、多丛芽诱导与增殖以及生根培养等最佳培养基配方。目前已将欧洲甜樱桃×中国樱桃、大石早生李×泰安巴旦水杏及凯特杏×总统李等一批核果类果树远缘杂种定植于露地 ,其中欧洲甜樱桃与中国樱桃的种间杂种是国内外首次获得