应用均匀设计研究电场刺激及培养因素对甘蓝叶原生质体分裂频率的影响

应用均匀设计法探讨了电场刺激及培养因素对甘蓝叶原生质体分裂频率的影响。结果表明 ,电场方向和强度强烈地影响细胞分裂频率。培养成分对细胞分裂频率均有不同程度的影响。均匀设计能筛选最优的植物原生质体培养条件或培养基。结合“回归 -通径”分析 ,有效地综合评判和研究了各培养成分对原生质体分裂频率的作用     

结构阻尼系统的一种子结构综合方法

本文对结构阻尼系统提出一种模态综合方法,该方法特别适合于部件对接界面自由度较少情况下的结构综合.给出了一个算例来说明方法的有效性.     

Activities of biotransformation enzymes in pheasant (Phasianus colchicus) and their modulation by in vivo administration of mebendazole and flubendazole

Basal activities of certain pheasant hepatic and intestinal biotransformation enzymes and modulation of their activities by anthelmintics flubendazole (FLBZ) and mebendazole (MBZ) were investigated in subcellular fractions that were prepared from liver and small intestine of control and FLBZ or MBZ treated birds. Several oxidation, reduction and conjugation enzyme activities were assessed. In the liver, treatment of pheasants by FLBZ or MBZ caused very slight or no changes in monooxygenase activities and conjugation enzymes. More significative changes were detected in small intestine. Metyrapone and daunorubicin reductase activities were increased by both substances in the liver. This is the first evidence that certain benzimidazoles modulate reductases of carbonyl group. With respect to the relatively slight extent of the changes caused by FLBZ or MBZ we can assume that repeated administration of therapeutic doses of both FLBZ and MBZ has probably no serious influence on pheasant biotransformation enzyme system.     

Effect of gonadotropin-releasing hormone analogue on human breast cancer cell line in vitro

AIM: To investigate the effects of gonadotropin-releasing hormone (GnRH) analogue on the growth of breast cancer cell lines MCF-7 and MDA-MB-231 in vitro and to explore the related mechanisms with PI3K/Akt or ERK/MAPK pathways. METHODS: The proliferation of human breast cancer cell lines MCF-7 and MDA-MB-231 treatment with triptorelin was detected by MTT assay and the distribution of the cell cycle was determined by flow cytometry. The phosphorylation of the ERK1/2 and Akt was evaluated by Western blotting. RESULTS: Triptorelin inhibited the proliferation of MCF-7 cells at concentration of 10^-5 mol/L after treated for 192 h or at concentration of 10^-4 mol/L after treated for 168 h and 192 h. Triptorelin inhibited the proliferation of MDA-MB-231 cells at concentration of 10^-4 mol/L after treated for 192 h (P<0.05).Treatment with triptorelin for 192 h at concentration of 10^-4 mol/L had no statistical significance effect on phosphorylation of ERK1/2 and Akt(P>0.05).CONCLUSION: Inhibitory effect of GnRH analogue triptorelin on human breast cancer cells is not just the connection with the down-regulation of pituitary hormone, but also a direct inhibitory effect. The role may not be involved in the activation of ERK/MAPK and PI3K/Akt signaling pathways.     

Role of kisspeptin/GPR54 signaling pathways in prompting formation of precocious puberty

AIM: To explore the role of kisspeptin/GPR54 signaling pathway in the pathogenesis of precocious puberty based on female precocious puberty rat model induced by the single dose of danazol. METHODS: Female SD rats aged 3 d were randomly divided into normal group, vehicle group and model group. On the 5th day, the model rats were given a single subcutaneous injection of danazol with ethanol and ethylene glycol mixture. All rats were executed on 15 d, 25 d, 30 d, 35 d and 40 d, and the samples were collected to observe the sexual organ development. The levels of E₂, FSH and LH in peripheral blood were measured by ELISA. The Kiss-1 mRNA expression of Kiss-1, GPR54 and GnRH in hypothalamus was detected by real-time PCR. The kisspeptin expression in rat hypothalamus was observed by immunofluorescence. RESULTS: The time for puberty onset and sexual maturation in the model rats was significantly earlier than that in normal group and vehicle group. On days 25 and 30, the levels of peripheral sex hormones and uterine coefficients in model group were significantly higher than those in normal group and vehicle group. On days 25 and 30, the ovarian morphological development in the model rats was significantly earlier than that in normal group. On day 25, the mRNA expression of hypothalamic Kiss-1 and GnRH, and kisspeptin expression of hypothalamic arcuate nucleus in the model rats significantly increased compared with normal group and vehicle group. On day 30, kisspeptin expression of hypothalamic arcuate nucleus in the model rats decreased compared with normal group and vehicle group. On day 35, the mRNA expression of Kiss-1 and GnRH in the model rats decreased compared with normal group and vehicle group. The mRNA expression of GPR54 had no obvious difference among all groups. CONCLUSION: The Kiss-1 mRNA and kisspeptin expression in the model rats with precocious puberty is significantly increased in the hypothalamus during onset of puberty, suggesting that kisspeptin is an initiating factor for precocious puberty. Kisspeptin/GPR54 signaling pathway may play an important role in the occurrence of precocious puberty.     

Development of cell model of polymer/liquid crystal and effect of their elasticity on adhesion of rBM-MSCs

AIM: To develop the cell model of polymer/liquid crystal and to study the effect of their elasticity on the adhesion of rat bone marrow mesenchymal stem cells (rBM-MSCs). METHODS: Using the method of solvent evaporation induced phase separation, the cell model of polymer/liquid crystal was constructed. The surface morphology and phase separation structure were determined by polarized optical microscopy (POM), scanning electron microscopy (SEM) and small angle X-ray scattering (SAXS). rBM-MSCs were separated and expanded by adherent culture. The surface markers of rBM-MSCs were detected by flow cytometry. The cells were induced to osteogenic differentiation and adipogenic differentiation for 2 weeks. After 3 passages, the cells were divided into 4 groups, including total PU control group, 10% membrane group, 30% membrane group and 50% membrane group. The cells were then incubated with rhodamine phalloidin for cytoskeleton staining and were observed under the confocal laser scanning microscope after cultured for 24 h. RESULTS: The cell model of polymer/liquid crystal was constructed successfully using the method of solvent evaporation induced phase separation. Flow cytometry results showed that the rBM-MSCs positively expressed CD29, CD44 and CD90, and negatively expressed CD34 and CD45. After stained with alizarin red S and oil red O, the calcium nodule and lipid droplets in rBM-MSCs were observed obviously. The cytoskeleton staining result indicated that the area in total PU control group, 10% membrane group and 30% membrane group were greater, and the actin microfilaments were also clearer than that in 50% membrane group. CONCLUSION: The cell model with suitable content of liquid crystal made a contribution to the rBM-MSCs' adhesion, but too much liquid crystal inhibits cell adhesion.     

Effects of maternal limb ischemic preconditioning on structural and functional changes of mitochondria in fetal hippocampal neurons induced by intrauterine distress-reoxygenation in rats

AIM: To investigate the effects of maternal limb ischemic preconditioning (LIP) on the mitochondrial structures and functions of the hippocampal neurons induced by reoxygenation in the intrauterine distress fetal rats. METHODS: Pregnant rats (n=40) were randomly divided into 4 groups: sham (S) group, LIP group, fetal distress (FD) group and LIP+FD group. Intrauterine ischemia model was established through the experimental design. The ultrastructure of the mitochondria in CA1 area of the hippocampus was observed. The mitochondrial membrane potential and reactive oxygen species (ROS) were measured. The content of ATP and MDA in the hippocampus tissue was detected. The activity of Mn-SOD was observed. RESULTS: Compared with sham group, the ultrastructure of mitochondria in CA1 area of the hippocampus was damaged in FD group and LIP+FD group. The mitochondrial membrane potential, the content of ATP and the activity of Mn-SOD were decreased. However, the content of ROS and MDA was increased. Compared with FD group, the ultrastructure of mitochondria in CA1 area of the hippocampus was intact in LIP+FD group. Furthermore, the reduced mitochondrial membrane potential and ATP content were inhibited. The activity of Mn-SOD was increased, but the content of ROS and MDA was decreased in LIP+FD group. CONCLUSION: Limb ischemia preconditioning inhibits the damage the mitochondria of fetal hippocampal neurons induced by reoxygenation in the intrauterine distress fetal rats.     

Proteomic study of myocardial mitochondria with ischemia/reperfusion injury and pinacidil postconditioning in isolated rat hearts

AIM: To investigate the protective effect of pinacidil postconditioning on rat myocardium suffering ischemia/reperfusion injury by mitochondrial proteomics. METHODS: Langendorff apparatus was used to establish the model of myocardial ischemia/reperfusion injury. Sprague-Dawley rats were randomly divided into 2 groups: pinacidil postconditioning group (Pina group) and ischemia/reperfusion injury group (I/R group). After 20 min of perfusion with K-H solution, the perfusion was suspended for 40-min (global ischemia) follow by 60 min of reperfusion in I/R group. In Pina group at the end of 40 min global ischemia, the isolated hearts were perfused with K-H solution containing pinacidil (50 μmol/L) for 2 min followed 58-min perfusion with regular K-H solution. Total proteins extracted from the mitochondria were applied to the two-dimensional gel electrophoresis (2-DE). The differentially expressed protein spots over 2 times were evaluated by a software. Then they were subjected to in-gel digestion, and analyzed by spectrometry. RESULTS: The expression levels of NDUFA10, NDUFS2 and NDUFV2 were elevated but those of IDHA and ECH1 were decreased in Pina group compared with I/R group. Interestingly, 2 spots in the 2-DE map were identified as ATPase subunit δ. The expression levels of one spot was elevated, while the other was decreased. CONCLUSION: Pinacidil postconditioning may decrease the degree of increased expression levels of NDUFA10, NDUFS2 and NDUFV2, promote the expression of IDHA and ECH1, and induce the phosphorylation of ATPase subunit δ, which may be related to the protective mechanism of pinacidil postconditioning.     

金银花和红银花挥发性成分的顶空固相微萃取#br# 气质联用检测与比较

 采用顶空固相微萃取气质联用（SPME–GC–MS）的方法检测红银花与金银花新鲜花蕾和 干花蕾的挥发性成分。4 种花蕾样品共鉴定出67 种挥发性物质,其中新鲜红银花与金银花分别检测出48 种和46 种,经烘干处理的干红银花与金银花分别检测出34 种和37 种。[3.2.1.0（1,5）]三环辛烷、β– 芳樟醇、5–甲基–2–己醇和以及十六烷是4 种样品共有的成分。其中红银花挥发性成分主要以烃类、 醇酮类为主,β–芳樟醇含量显著高于金银花;样品在由新鲜到烘干这一过程中,烃类和酯类挥发性物质 含量降低,醛类和醇酮类挥发性物质显著增加。     

不同菜豆品种4种抗营养因子水平差异分析

菜豆中植物凝集素、皂苷、胰蛋白酶抑制剂和植酸等抗营养因子是引起菜豆食物中毒的内源物。以56份食荚菜豆品种为材料,测定了鲜荚中4种抗营养因子含量或活性。结果表明,供试菜豆品种群体4种抗营养因子水平均存在极显著差异。植物凝集素含量(均值为1.743 mg·g~(-1))和胰蛋白酶抑制剂活性(均值为1.680 mg·g~(-1))变异系数均在100%以上,品种频率分布曲线主峰明显,但在极高和极低区域均有品种间断分布,出现了品种水平的数量等级差异;皂苷含量(均值为3.730 mg·g~(-1))和植酸含量(均值为3.102 mg·g~(-1))变异系数均低于41%,品种频率分布曲线呈近正态分布,主峰明显,双尾有低频率连续分布。相关分析表明,植物凝集素含量与胰蛋白酶抑制剂活性呈极显著正相关。聚类分析将供试56份菜豆品种划分为3个品种群,近80%品种聚于抗营养因子中等水平品种群,近12%品种居于较低水平。由于菜豆植物凝集素是食用致毒性的主要内源物,同时其和胰蛋白酶抑制剂等均与菜豆田间抗病虫性密切相关,预示着筛选出极低水平植物凝集素的菜豆品种是可行的,但同时可能会降低其田间抗病虫性。