Role of 17-AAG in inducing apoptosis and cell cycle arrest of HCT-15 cells

AIM: To investigate the effects of 17-AAG on apoptosis and cell cycle of HCT-15 cells and to clarify the related mechanisms. METHODS: MTT method was employed to evaluate the inhibitory effects of 17-AAG with Aifferent time and different doses on the proliferation of HCT-15 cells. The cells were stained with Annexin V-FITC/propidiumiodide and measured by flow cytometry. The expression of STAT3, cyclin D1, Cyt C, caspase 9 and caspase 3 at mRNA and protein levels was determined by RT-PCR and Western blotting. RESULTS: Treatment with 17-AAG at concentration of 1.25~20 mg/L for 24 h and 48 h significantly inhibited the activity of HCT-15 cells at both time-and concentration-dependent manners. Treatment with 17-AAG at concentrations of 0.425, 0.85 and 1.7 mg/L for 48 h significantly induced apoptosis and cell cycle arrest of HCT-15 cells. The exposure of 17-AAG at concentrations of 0.425, 0.85 and 1.7 mg/L for 48 h to the HCT-15 cells significantly down-regulated the expression of STAT3 and cyclin D1 at mRNA and protein levels, but up-regulated Cyt C, caspase 9 and caspase 3 mRNA and protein in a concentration-dependent manner. CONCLUSION: 17-AAG inhibits the cell activity, induces apoptosis and G₁ arrest by down-regulating the expression of cyclin D1, and promoting the mitochondria apoptosis through STAT3 pathway.     

Relationship between Th17 cells infiltrated in lungs and pulmonary vascular reconstruction under hypobarric hypoxia

AIM: To investigate the changes of retinoid-related orphan receptor γt(RORγt) mRNA and interleukin-17(IL-17) protein in the lung tissue under hypobaric hypoxia, and the relationship between Th17 cells and hypoxic pulmonary vascular reconstruction. METHODS: Male BALB/c mice(n=50) were randomly divided into control group and 3 d, 7 d, 14 d and 28 d hypobaric hypoxia groups. The mice in hypobaric hypoxia groups were housed in a hypobaric hypoxia chamber(simulated altitude of 6 000 m) for 3 d, 7 d, 14 d or 28 d. The mice in control group were housed in normal pressure and oxygen environment. The hemodynamic data were recorded by cardiac catheterization. The hypertrophy of right ventricle was evaluated by the ratio of weight of the right ventricle to the weight of the left ventricle plus interventri-cular septum, and the right ventricular weight over body weight. The spleen was collected and the proportions of the Th17(CD4⁺IL-17⁺RORγt⁺) cells were detected by flow cytometry. The serum levels of IL-4, IL-6 and IL-17 and the change of IL-17 in the lung tissue were measured by ELISA. The mRNA expression of RORγt in the spleen and lung tissues was measured by RT-qPCR. RESULTS: Compared with control group, the mouse right ventricular systolic pressure, the hypertrophy index of right ventricle and the serum IL-17 level were significantly elevated in hypoxia groups, which was consistent with the results of flow cytometry. The mRNA expression of RORγt in the lung tissue was also significantly increased in 7 d, 14 d and 28 d hypoxia groups. The expression of IL-17 in the lung tissue was significantly increased in 14 d and 28 d hypoxia groups. CONCLUSION: Hypoxia promotes differentiation of Th0 cells to Th17 cells in the spleen. The Th17 cells infiltrated in the lung tissue under hypobarric hypoxia are involved in pulmonary vascular reconstruction.     

甬优17作烟后稻机插高产栽培技术

通过近2 a对甬优17作烟后稻机插栽培试验示范观察,总结了适时播种,培育壮秧,合理密植,科学管理肥水和防治病虫害等高产栽培配套技术,以期为"春烟—晚稻"模式种植区的粮食增产提供参考。     

Little effect of forest age on oribatid mites on the bark of trees

This study investigates the effect of forest age (20-, 50-, 160-year-old, and primeval forest) on oribatid mite communities on the bark of oak trees in the National Park Pusza Białowieża in eastern Poland. We hypothesized that oribatid mite diversity on bark peaks at forests of intermediate age and that the number of parthenogenetic species of oribatid mites is highest in young stands. In contrast to these hypotheses, the diversity, density, number of juveniles, community structure and the mode of reproduction of oribatid mite species were not significantly affected by forest age. None of the oribatid mite species occurred exclusively on trees of a specific age. The results suggest that oribatid mite communities on the bark of trees are minimally affected by tree harvesting regimes. In contrast to oribatid mites in soil, communities on bark appear to be less sensitive to disturbances.     

西农萨能奶山羊母羔初情期的研究

对30只西农萨能奶山羊母羔的初情期体重、日龄、发情表现,发情周期及17β—雌二醇(17β-E_2)和孕酮(P_4)水平变化进行了研究。研究结果表明:1.初情期体重和日龄分别为10.2(8.0—12.9)kg和54.7(43—71)天,初情期体重和日龄的相关系数为0.798,初情期日龄和出生日期的相关系数则为0.963;2.第一个发情周期平均为8.3(2—37)天,其中短周期(2—8天)羊占62.4％;3.发情表现的特点是阴道和阴门下角有灰白色粘液;4.初情期17β—E_2(pg/ml)/P_4(ng/ml)比值为141.1,17β-E_2和P_4的分泌关系有同步、反步和部分同步部分反步现象。     

Breast cancer stem cells cultured without insulin and effect of estrogen induction

AIM:To determine whether suspension culture medium without insulin can be used to feed breast cancer tumorsphere, or not. METHODS:MCF7 cells were used to build tumorsphere. The morphological changes, CD44+ CD24- expression, aldehyde dehydrogenase 1 (ALDH1) expression and multiple division ability were measured to identify the breast cancer stem cells and to detect the function of 17β-estradiol (E2β) in tumorsphere of MCF7 cells. RESULTS:The tumorshere, each containing 30 to 60 cells, was obtained by the method of insulin-removal suspension culture. These cells were cytokeratin 18 and CD10 proteins positive, and the number of CD44+ CD24- cells and ALDH1 protein expression were significantly higher than the adherent cultured cells (P<0.05). Using 10-10 mol/L E2β to treat the tumorshere for 7 d, the tumor cell number and volume were significantly increased. Using 10-10 mol/L E2β to treat the tumorshere for 24 h, the CD44+ CD24-cells and ALDH1 protein expression were significantly higher than those in non-treatment group (P<0.05). CONCLUSION: Suspension culture medium without insulin can be used to feed breast cancer tumorsphere. These tumorsphere could be used as a model to determine the function of E2β in breast cancer stem cell research.     

17β-雌二醇对AC细胞活性、细胞周期和超微结构的影响

通过免疫荧光获得雌激素α、β受体在体外星形胶质细胞（Acrocyte,AC）中表达的形态学证据;CCK-8法获得脂多糖（Lipopolysaccharide,LPS）激发AC的适宜浓度。在此基础上,研究了17β-雌二醇（17β-estradiol,E2）对AC的细胞活性、细胞周期和超微结构的影响。结果显示：E2作用AC 48h,10nmol/L E2可提高AC细胞活性和G2＋S%（P〈0.05）,而100,1 000nmol/L E2可降低AC细胞活性（P〈0.05）,100nmol/L E2还可降低G2＋S%（P〈0.05）;100nmol/L E2作用AC 24,48,72h,均可降低AC细胞活性和G2＋S%,以作用48h时抑制作用最强（P〈0.05）;10nmol/L E2可引起AC线粒体数量增加,细胞核分裂增多,但线粒体、粗面内质网结构正常,而100nmol/L E2可导致AC线粒体肿胀或空泡化,粗面内质网扩张或断裂。结果表明,低浓度（10nmol/L）E2促进AC增殖;高浓度（100,1 000nmol/L）E2抑制AC增殖。     

蒙古马突触融合蛋白17(STX17)基因多态性及在不同毛色皮肤组织中的表达分析

为研究突触融合蛋白17(STX17)基因与蒙古马毛色之间的相关性及其作用机制.本研究利用半巢式PCR技术及实时荧光定量PCR技术对蒙古马STX17基因第6内含子的多态性以及不同毛色蒙古马皮肤组织中STX17基因的相对表达量进行了检测.巢式PCR结果显示:STX17基因在青毛蒙古马第6内含子处存在4.6 kb的重复片段,在其它毛色中不存在该重复片段;荧光定量PCR结果显示:STX17基因在蒙古马青毛皮肤组织中表达量最高,在骝毛皮肤组织中的表达量最低.结果证明蒙古马STX17基因与其青毛表型具有典型的相关性.     

不同浓度雌二醇对猪卵母细胞体外成熟和孤雌激活胚胎的影响

为了探讨雌二醇（17β-estrodiol,E2）对猪卵母细胞体外成熟及孤雌激活后胚胎早期发育的影响,在卵母细胞体外成熟培养基中添加不同浓度雌二醇,研究卵裂率和囊胚率的变化。以未添加雌二醇的基础液为对照组,比较分析各组卵母细胞核成熟效率、孤雌激活后胚胎的卵裂率、囊胚发育率。结果表明,成熟液中添加1μg／mL雌二醇（E2）对猪卵母细胞的体外成熟具有明显的促进作用,而添加100μg／mL雌二醇（E2）对猪卵母细胞体外成熟具有明显的抑制作用。