Adhesion of bone marrow stromal cell in children with acute lymphoblastic leukemia

AIM and METHOD: The adhesion behavior of bone marrow stromal cells(BMSC) in children with acute lymphoblastic leukemia(ALL) to bone marrow mononuclear cells(BMMC) and Raji cells were investigated by MTT method. The expression of adhesion molecules ICAM-1 and VCAM-1 were detected by flow cytometry. RESULTS: The adhesion ability of BMSC in acute period of ALL to BMMC was lower than that of control group. The adhesion ability of BMSC in long term remission period of ALL to Raji cells higher than that of control group. The expression of ICAM-1 on the surface of BMSC in acute period of ALL is much lower than that of control group. CONCLUSIONS: The adhesion of BMSC to BMMC or tumor cells in children with ALL was abnormal. The abnormal adhesion behavior might be partially due to the changed expression of adhesion molecules on BMSC.     

Induction of stem cells from adult Beagle canine bone marrow into chondrocytes in vitro

AIM：To explore the methods of the differentiation from adult Beagle canine bone marrow stem cells (BMSCs) into chondrocytes in vitro and determine the factors involved in the differentiation process.METHODS：About 10 mL BMSCs were aspirated from canine femoral bone,primarily cultured and subcultured in vitro.TGF-β1 was added into the culture medium.BMSCs were cultured and expanded in the medium until they reached the required quantity.BMSCs were induced to differentiate into chondrocytes at high cell density.Matrix of cartilage cells was detected by toludine blue stain,and cartilage specific collagen Ⅱ was detected by immunohistochemistry.RESULTS：The structure of cellular cartilage form BMSCs was uniformly positive of toludine blue staining.Immunohistochemical staining was positive for the collagenⅡ.CONCLUSION：Application of TGF-β1 may induce canine bone marrow stem cells into chondrocytes in vitro,which can be used as seeding cells in cartilage tissue engineering.     

Calpain regulates signal transduction contributing to cardiomyocyte hypertrophy

AIM: To induce rat bone marrow mesenchymal stem cells (BMSC) into cardiomyocytes and investigate the influence of serum coming from acute myocardial infarction (AMI) rat on the procedure. METHODS: The passage 3 BMSC were divided into six groups: groupⅠwas control group; groupⅡwas induced with 5-azacytidine; group Ⅲ was induced with 5-azacytidine and serum from AMI rat; group Ⅳ was induced with 5-azacytidine and serum from normal rat; group V and group Ⅵ were induced with serum from AMI rat or normal rat. The cardiac troponin T, GATA-4 and desmin were detected 30 days after induction. RESULTS: After inducing by 5-azacytidine, 5-azacytidine and two kinds of serum, some cells in the three groups differentiated into cardiac like cells. The expressions of cardiac troponin T, GATA-4 and desmin were positive in cells differentiated from BMSC. The troponin T expression in control group and group inducing by AMI serum alone were negative but GATA-4 and desmin expressed weakly. Some cells induced with 5-azacytidine and serum were slowly beating 2 weeks after induction, but the cells induced with 5-azacytidine alone was not beating.CONCLUSION: Serum from AMI can not induce BMSC to differentiate into cardiomyocytes, but it promotes BMSC differentiate into cardiomyocytes induced by 5-azacytidine and facilitate the differentiated cells to mature.     

Effects of bFGF and insulin on proliferation of mice cartilage tissue cells

AIM:To explore the effects of basic fibroblast growth factor ( bFGF) and insulin on the cell proliferation and differentiation in primary cartilage cells. METHODS:After induction with different concentrations of bFGF and insulin, cell proliferation was measured with WST-1 method and fluoroscope methods. RESULTS:bFGF and insulin exerted their related action on primary cartilage cells in 0.4% fatal bovine serum at different concentrations. 25 μg/L bFGF and 5 mg/L insulin promoted cell proliferation significantly. CONCLUSION:bFGF and insulin prolong the survival time and promote cell proliferation in primary cartilage cells.     

Molecular mechanism of BMSC intracerebral transplantation in improving learning and memory abilities of AD mice

AIM: To investigate the effect of bone marrow mesenchymal stem cell (BMSC) transplantation on learning and memory abilities and pathological changes of Alzheimer disease (AD) mice and the molecular mechanisms. METHODS: C57/BL6 wild-type (WT) and transgenic (Tg) mice were randomly divided into 4 groups:WT/PBS group, WT/BMSCs group, Tg/PBS group and Tg/BMSCs group. The mice were administered with PBS or BMSCs via intracerebroventricular injection. Spatial learning and memory abilities of the mice were evaluated by Morris water maze test on the 3rd day after surgery. Real-time PCR was applied to detect the mRNA expression of CX3C chemokine ligand 1 (CX3CL1), CX3C chemokine receptor 1 (CX3CR1), IL-1β, TNF-α, Nurr1, YM1, insulin-degrading enzyme (IDE) and matrix metalloproteinase 9 (MMP9). The protein levels of CX3CL1 and Aβ42 were measured by ELISA. Western blot was used to detect the protein expression of postsynaptic density protein 95 (PSD95) and synaptophysin (SYP). RESULTS: The transplanted BMSCs were observed near the hippocampus of APP/PS1 mice on the 10th postoperative day. The escape latency of the mice in Tg/PBS group was significantly longer than that in the WT/PBS mice (P<0.05). Compared with Tg/PBS group, the escape latency of Tg/BMSCs group was significantly shorter (P<0.05), and the mRNA and protein levels of CX3CL1 in Tg/BMSCs group were significantly higher than those in Tg/PBS group (P<0.01). The results of immunohistofluorescence staining showed that BMSC transplantation promoted the activation of microglia in the brain of WT and Tg mice. The mRNA expression of YM1 was up-regulated in WT/BMSCs group and Tg/BMSCs group (P<0.05). Compared with WT/PBS mice, the mRNA expression of TNF-α in the cortex and hippocampus of Tg/PBS group was significantly increased (P<0.05), and the mRNA expression of Nurr1 in the cortex was significantly decreased (P<0.01). Meanwhile, the mRNA expression of TNF-α in the cortex of Tg/BMSCs mice was decreased (P<0.01) and the mRNA expression of CX3CR1 and Nurr1 was up-regulated compared with Tg/PBS group (P<0.05). The results of Western blot showed that the protein levels of PSD95, p85, p110 and p-Akt in Tg/BMSCs group were significantly higher than those in Tg/PBS group (P<0.05). Finally, BMSC transplantation reduced the protein level of Aβ42 in APP/PS1 mice (P<0.05), and increased the mRNA expression of IDE and MMP9 in the hippocampus (P<0.05). CONCLUSION: BMSC transplantation modulates neuroinflammatory responses and promotes neuroprotective factor and synaptic protein expression, thus improving the learning and memory abilities in the APP/PS1 mice, which may be achieved by up-regulating the expression of CX3CL1.     

基因工程雄性不育及辣椒杂种优势利用

综述了我国近年来辣椒杂种优势利用的现状及其研究进展,分析了目前杂交制种中的技术弊端,着重探讨基因工程在辣椒杂种优势上的利用,介绍应用基因工程创造辣椒雄性不育系原理和方法,以及提出基因工程雄性不育的保持和育性恢复途径。     

植物细胞工程在菊花育种中的应用

综述了近几年来细胞工程应用于菊花辐射诱变、化学诱变、体细胞杂交和转基因育种方面研究进展,其中详细介绍了采用拯救性茎尖培养获得类病毒脱毒苗的研究,和薄层细胞培养减少转基因植株的嵌合体、提高转化率方面的研究。     

葱蒜类蔬菜种质资源创新研究进展

综述了葱蒜类蔬菜种质资源创新的研究进展。创新手段主要有远缘杂交、细胞工程、基因工程等；创造了许多新的重要性状，如抗病虫、抗除草剂、耐盐、雄性不育等；并对我国葱蒜类蔬菜种质资源创新的应用前景进行了讨论。     

Time window of bone marrow stem cell homing to ischemia myocardium after acute myocardial infarction in rats

AIM: To investigate the time window of bone marrow stem cell homing to the inchemia myocardium. METHODS: 110 SD male rats were randomly divided into four groups: GM-CSF-treated group (n=40) and sham-treated group (n=15) were given GM-CSF (50 μg/kg/d) for 5days. The control group (n=40) and its sham-treated group (n=15) was injected equal volume of saline, acute myocardiar infarction were induced by LAD ligation in each group. At 1, 3, 5, 10 days in each group, the homed bone marrow stem cells were detected by expression of c-kit with immunohistochemical methods. Cardiac performance and pathological changes were examined at 28 days. RESULTS: At 28 days, both systolic function and diastolic function in GM-CSF-treated group were significantly higher than those in control group (P<0.05). Histological observation indicated that the tissue repair in GM-CSF-treated group was significantly higher than that in control group (P<0.05). The GM-CSF-treated group and control group both had the homed c-kit+ cells at 1, 3 and 5 days. At 5 days, both had high number of c-kit+ cells, but the homed c－kit+ cells in GM-CSF-treated group were significantly higher than that in control group (P<0.05) at 1, 3 and 5 days (P<0.05). At 10 days both group had no new homed c-kit+ cells. CONCLUSION: The time window of BMSC homing is within 10 days after AMI in rats.     

高职园林工程专业人才培养目标和教学改革探讨——以成都农业科技职业学院园林工程技术专业为例

以成都农业科技职业学院为例,浅析了我院园林工程技术专业人才培养目标是培养德、智、体、美全面发展,适应社会竞争需求,以"四条能力"主线为核心,能从事园林工程规划设计、园林工程施工、园林工程项目管理及园林植物栽植与养护的应用型高级技术人才。围绕这一目标,从课程设置、教学方法、教学条件、师资培养等几个方面对高职学院园林工程技术专业人才培养和教学改革进行了探讨。     

启动子克隆概述

启动子是基因表达调控的重要顺式元件，也是基因工程表达载体的一个重要元件。笔者阐述了启动子的结构、分类、克隆方法及其意义，并介绍了目前食用菌中已分离到的启动子。     

Effects of Qiancaofang fumigation on expression of YAP/Smad3 in rats with knee osteoarthritis

AIM: To observe the effects of Qiancaofang fumigation on the expression of Yes-associated protein (YAP) and Smad3 in the cartilage of the rats with knee osteoarthritis (KOA), and to investigate the protective effects and mechanisms of Qiancaofang fumigation on the cartilage of the rats with KOA. METHODS: SD rats (n=32) were randomly divided into 4 groups:control group, model group, Chinese medicine group and hyaluronic acid (HA) group. The improved Hulth method was used to construct the rat model of KOA in model group, Chinese medicine group and HA group. The rats in Chinese medicine group were treated with Qiancaofang fumigation. The rats in HA group were treated with intra-articular injection of HA. After treatments, the cartilage tissues and serum of the rats in each group were taken. Hematoxylin-eosin (HE) staining and Mankin's scores were measured. The expression levels of cartilage oligomefic matrix protein (COMP) and C-telopeptide of type Ⅱ collagen (CTX-Ⅱ) in the serum were detected. The method of TdT-mediated dUTP nick-end labeling (TUNEL) was used to observe the apoptosis of the cartilage cells. Immunohistochemistry was performed to detect the expression of YAP and high mobility group protein B2 (HMGB2). Western blot was used to detect the expression of YAP, Smad3, bone morphogenetic protein 2 (BMP2), Bax and Bcl-2. RESULTS: The results of HE staining showed that the cartilage cells in model group were disordered and the structure was not clear. Compared with model group, the cartilage surface in Chinese medicine group was more smooth, the structure was clearer, and the staining of the cartilage matrix was more uniform. Compared with model group, the Mankin's scores in Chinese medicine group and HA group were significantly decreased (P<0.05), the serum concentration of CTX-Ⅱ was significantly increased, the serum concentration of COMP was significantly decreased, and the apoptosis rate of the cells was decreased (P<0.05). The results of immunohistochemistry and Western blot showed that the expression levels of YAP, HMGB2, Smad3 and Bcl-2 were increased in Chinese medicine group and HA group, and the expression levels of BMP2 and Bax were decreased as compared with model group (P<0.05). CONCLUSION: Qiancaofang fumigation relieves KOA partially through the YAP/Smad3 signaling pathway.     

Expression of Akt and ERK1/2 in human osteoarthritic chondrocytes

AIM: To investigate the expression of protein kinase B (Akt) and extracellular signal-regulated kinase 1/2 (ERK1/2) in normal and osteoarthritic chondrocytes. METHODS: The samples of knee cartilage were obtained from the normal donors (n=5) and the patients (n=18) undergoing total knee arthroplasty with the diagnosis of osteoarthritis (OA). The expression of p-Akt and p- ERK1/2 in the normal and osteoarthritic cartilage tissues was detected by the method of immunohistochemistry. The chondrocytes were isolated and identified by toluidine blue staining and immunohistochemical method. The expression levels of Akt, p-Akt, ERK1/2,p-ERK1/2,phosphorylated 70-kD ribosomal protein S6 kinase(p-p70S6K) and proliferating cell nuclear antigen(PCNA) were tested in normal and osteoarthritic chondrocytes by Western blotting. Real-time fluorescence quantitative PCR was used to measured the expression levels of aggrecan and type II collagen gene in normal and osteoarthritic chondrocytes. RESULTS: The expression of p-Akt in normal cartilage was higher than that in OA cartilage. The expression of p- ERK1/2 in OA cartilage was higher than that in normal cartilage. Compared with the normal chondrocytes, the expression of p-Akt and p-p70S6K, and the mRNA levels of aggrecan and type II collagen were increased (P<0.05), and the expression of p-ERK1/2 and PCNA was decreased in OA chondrocytes (P<0.05). CONCLUSION: Akt might regulate aggrecan and type II collagen synthesis via p-p70S6K, and ERK1/2 might regulate OA chondrocyte proliferation through PCNA. Both Akt and ERK1/2 play important roles in the pathogenesis of OA.     

岩质边坡植被护坡技术在深圳的应用研究

在自然环境条件下,岩体(特别是硬岩)裸露难以依靠自然恢复岩面生态,而人工干预是岩质边坡生态恢复的有效技术途径。基于生态护坡技术和植被群落构建思想,根据边坡修复中采用不同的生态护坡技术和植物物种选择而呈现出多样的恢复效果,从边坡特点与护坡技术、植生基材设计与配比、植物配置和植被群落构建的关系等方面,对深圳岩质边坡人工生态恢复工程案例进行研究,以期对深化深圳边坡生态恢复工作、创新边坡生态恢复工程技术、提高工程设计水平具有促进作用;同时,也可以进一步丰富边坡植被恢复理论,完善边坡修复工程的实践经验。     

高等真菌基因工程研究进展

本文综述了高等真菌遗传转化所采用的选择标记和外源DNA导入受体真菌的几种方法,比较了高等真菌复制型转化与整合型转化的转化子的遗传稳定性,简述了高等真菌基因工程在工农业生产中的应用现状,展望了基因工程技术在食用蕈菌遗传育种中的应用前景。     

Effect of TNF-α on the cartilage endplate chondrocytes of New Zealand white rabbits

AIM:To evaluate the biological roles of TNF-α on the cartilage endplate cells (chondrocytes). METHODS:The chondrocytes were isolated and harvested from the cartilage endplate of New Zealand rabbits and then the biological characteristics of cells were identified by methods such as toluidine blue staining for type Ⅱ collagen. After different concentrations of TNF-α were added to culture medium respectively, the rate of the proliferation of chondrocytes in different time was measured with MTT. The protein expressions of Bax, Bcl-2, Fas and caspase-3 were measured by immunocytochemistry. The changes of the mRNA of aggrecan and type Ⅱ collagen were measured by RT-PCR. RESULTS:The TNF-α at concentration of 50 μg/L and 100 μg/L decreased the rate of the proliferation on chondrocytes. Though TNF-α at concentrations of 10 μg/L and 50 μg/L increased the level of Bax, Fas and caspase-3, only 50 μg/L TNF-α decreased the level of Bcl-2. TNF-α at concentrations of 10 μg/L and 50 μg/L decreased the level of collagen IIa mRNA and only 50 μg/L TNF-α decreased the level of aggrecan. CONCLUSION:TNF-α not only inhibits the proliferation and the matrix synthesis in chondrocytes, but also increases the expression of pro-apoptotic factors in chondrocytes.     

基因工程与猕猴桃种质改良

利用植物基因工程进行猕猴桃遗传改良潜力很大,目前已建立有效的基因转化体系,并获得了转基因植株。猕猴桃转基因主要包括果实发育、成熟与衰老及其抗逆性等有关的目的基因。总结了猕猴桃目的基因分离与克隆、基因转化方法和影响因素以及基因工程在猕猴桃种质改良中的应用等方面的研究进展,并对猕猴桃基因工程研究中存在的主要问题及应用前景进行了讨论。     

Differentiation of bone marrow mesenchymal stem cells into chondrocytes

AIM: To investigate the differentiation of human bone marrow mesenchymal stem cells (MSC) into chondrocytes in vitro and determine factors involving in the differentiation process. METHODS: MSC were separated from iliac bone marrow with lymphocyte separating medium using density centrifugation. Cells were cultured and expanded in medium until reaching required number. MSC was induced to differentiate into chondrocytes by adopting high cell density, supplying growth factor and using micromass culture. Cells were observed by HE staining. Matrix of cartilage was detected by alcian blue and toludine blue and cartilage specific collagen II was detected by immunohistochemistry. RESULTS: The structure of the micromass assumed that of cellular cartilage, alcian blue staining were uniformly positive and toludine blue detected diffuse metachromasia substance, cells uniformly expressed collagen Ⅱ. CONCLUSION: High cell density, growth factor and appropriate culture conditions are critical to induce differentiation of MSC into chondrocytes.