羊栖菜多糖诱导lovo细胞凋亡及其机理探讨

本课题研究羊栖菜多糖(Sargassum Fusiforme Polysaccharides,SFPS)诱导人大肠癌lovo细胞凋亡及凋亡过程中Caspase-3的变化及其意义。MTT法检测SFPS对大肠癌细胞增殖的抑制率;通过电镜、琼脂糖凝胶电泳、流式细胞术鉴定细胞凋亡;应用Western blot法测定Caspase-3酶原的变化; RT-PCR检测Caspase-3 mRNA表达。结果显示:SFPS作用lovo细胞24,36,48和72h的IC_(50)分别为375,355,178和60mg/L,表明对lovo细胞具有显著生长抑制作用。在电镜下,可见明显的细胞凋亡特征:细胞膜表面微绒毛减少、染色质固缩、边集,凋亡小体形成。在琼脂糖凝胶电泳中,药物浓度为5-300mg/L作用24h后,显示有凋亡细胞特有的DNA梯状条带;而500mg/L处理后梯状条带模糊,开始出现“涂片状”,表明在高药物浓度的作用下,细胞有坏死。流式细胞仪测得细胞凋亡率有剂量的依赖性;DNA直方图出现亚G1峰,但细胞周期时相的分布无明显改变。SFPS处理lovo细胞后,发现Caspase-3酶原蛋白表达降低,Caspase-3的mRNA高表达,并具有剂量和时间的依赖性。实验结果提示,SFPS在体外能够诱导lovo细胞凋亡,这可能是SFPS抑制肿瘤增殖的机制之一,而Caspase-3的活化参与了SFPS诱导lovo细胞凋亡的调控。     

油松种子直流电场生物学效应的研究

用５００ｋＶ／ｍ直流电场处理油松种子１０ｍｉｎ后,其发芽率和活力指数明显提高,浸水种子分别为对照的１２３．２％和１６１．１％;干种子分别为对照的１２４．９％和１３２．６％。通过对油松种子萌发过程有关生理生化指标的测定表明,直流电场处理可促进细胞膜的修复提高种子萌发阶段的生物氧化水平,加速新陈代谢,利于种子萌发和幼苗生长。     

Sister chromatid differential staining and NOR activity of BrdU-resistant sublines

Summary Two 30 g/ml BrdU-resistant sublines and two 60 g/ml BrdU-resistant sublines are induced from a Chinese hamster cell line Wg3h (HGPRT^–) by one-step and two-step selections, respectively. By inoculating the cells into BrdU-free medium or by adding more BrdU into the culture medium for 26–27 h, it was found that the two BrdU-resistant sublines analysed have very clear sister chromatid differential (SCD) staining patterns. This indicates that some of the nuclear DNA of the BrdU-resistant cells incorporate with BrdU to reach a kinetic balance. Frequencies of sister chromatid exchange (SCE) of the resistant cells are twice to four times as high as those of the Wg3h cells, depending on which BrdU-resistant subline is analysed. The SCE frequencies of the resistant cells also increase with the BrdU concentration in the medium. Analysis of silver-stained nucleolar organizer regions (NORs) indicates that the NOR activity of three out of the four BrdU-resistant sublines is significantly suppressed, i.e., averages of the Ag-NOR number and number of the chromosomes bearing Ag-NORs per cell decrease significantly. The degree of suppression for different BrdU-resistant sublines may be quite different. The suppressed NOR activity of the resistant cells can gradually be restored when the cells are inoculated into BrdU-free medium, but the recovery speed is far lower than that of the Wg3h cells. The suppression of the NOR activity of the BrdU-resistant sublines should be due to BrdU toxicity.     

Effect of density on larval development and female productivity ofTisbe holothuriae (Copepoda,Harpacticoida) under laboratory conditions

The harpacticoid copepodTisbe holothuriae has been cultivated in the Helgoland laboratory for more than 20 years. The effects of density on the larval development and the female productivity were studied by comparing two culture systems: (1) enclosed system, and (2) running-water system. In both systems, a nutritious mixed diet ofDunaliella tertiolecta, Skeletonema costatum, and granulatedMytilus edulis was offered. Larval mortality, larval development and female productivity are found to be significantly dependent on both the population density and specificity of the culture system. Increasing density causes higher larval mortality, longer larval development time, and a reduction in female productivity. In comparison with the enclosed system, the running-water system shows decisive advantages: larval mortality is about 20% lower, the rate of larval development is about two days shorter, and there is a very high rate of nauplii production. The sex ratio exhibits high variations, but in general, there is no clear relationship between sex ratio and population density. Nevertheless, when reared in the running-water system, a relatively high percentage of females (>45%) was found at lower densities.     

安徽烟草病毒类型及优势种初步研究

1978年,烟草病毒病在安徽烟区流行,导致凤阳县烟叶总产损失92.3％,引起了普遍震惊。1981—1984年作者对来自16个县、市552个病毒材料,经生物测定、血清反应、电镜观察,初步分离出CMV、TMV、PVY和PVX四种病毒。它们分别约占检测总数的82.79％、4.53％、2.54％和0.36％,其中CMV与长期视为优势种的TMV比值为18.3,除此,尚有约占检测总数9.8％的CMV和TMV、CMV和PVY复合侵染,以及不明类型的毒株。通过对田间烟草以及其他植物花叶病株的实际检测,进一步表明:CMV在烟区分布范围极广、出现频次最多,已形成了复杂的循环侵染系统,成为近期内烟草病毒病持续流行危害的首要毒原。     

AM真菌在草原生态系统中的功能

AM真菌是土壤生态系统中重要的微生物类群,能与陆地生态系统中80%以上的高等植物建立共生体系。目前,AM真菌在维持草原生态系统稳定性中的功能已经成为生态学研究的热点问题之一。基于此,从植物个体、种群、群落和生态系统等不同层次探究AM真菌在维持植物群落多样性和草原生态系统稳定性中的功能。分析发现在个体水平上,AM真菌对宿主植物具有促生效应、抑制效应或中性效应。在种群水平上,分析AM真菌对不同宿主植物吸收土壤矿质营养的分配和调控策略,围绕构成草原植被的两大组成成分:牧草和有毒植物,论述AM真菌对植物种群增长和衰败的调控机制,并从草原植物群落的物种多样性和稳定性角度,探讨AM真菌与植物群落之间的相关性。在生态系统水平上,围绕AM真菌对草原生态系统的演替和退化草原的修复等展开论述,以期为利用AM真菌开展草原生态系统保护和恢复治理提供理论依据,并对草原菌根生态学领域未来的研究进行展望。     

抗生素诱导的呼吸道菌群缺失对小鼠RSV感染的影响

目的 探究抗生素雾化暴露引起的呼吸道菌群缺失对小鼠呼吸道合胞病毒(RSV)感染的影响,为临床合理使用抗生素提供指导意见。 方法 32只BALB/c小鼠分为2组:雾化ddH2O对照组和雾化ABX组合抗生素组,处理6 d后,进行细菌16S rRNA基因PCR检测,构建呼吸道菌群缺失小鼠模型。上述2组组内再随机分为2小组,即PBS对照组(ddH2O+PBS,ABX+PBS)和RSV感染组(ddH2O+RSV,ABX+RSV),饲养至第14天。检测和分析各组小鼠支气管肺泡灌洗液(BALF)中的炎症细胞和相关细胞因子(TNFα、IL8、IL10及MCP1)的数量和水平,观察肺组织病理学状况及检测病毒载量。 结果 BALF中细菌DNA提取和16S rRNA基因PCR检测显示,雾化ABX组合抗生素处理能够有效地剔除呼吸道菌群。BALF中炎症细胞和相关细胞因子检测显示,ABX+RSV组炎症细胞总数明显增多(P2O+RSV组和ABX+RSV组小鼠肺部损伤明显加重(均P≤0.01),与ddH2O+RSV组相比较,ABX+RSV组的病毒载量明显升高(t=2.716 0,P=0.021 7)。 结论 雾化ABX组合抗生素不仅能够有效地剔除呼吸道菌群,而且明显增加了小鼠感染RSV的风险,导致呼吸道炎症加重,以及病毒载量升高。     

Hepatic Parenchymal Changes following Transcatheter Embolization and Chemoembolization in a Rabbit Tumor Model

Objective

To compare the effects of transcatheter arterial chemoembolization (TACE) with transcatheter arterial embolization (TAE) on liver function, hepatic damage, and hepatic fibrogenesis in a rabbit tumor model.

Materials and Methods

Thirty-nine New Zealand white rabbits implanted with VX2 tumors in the left liver lobes were randomly divided into three groups: TAE, TACE, and control group. In the TAE group (n = 15), polyvinyl alcohol particles (PVAs) were used for left hepatic artery embolization. In the TACE group (n = 15), the tumors were treated with left hepatic arterial infusions of a suspension of 10-hydroxycamptothecin and lipiodol, followed by embolization with PVAs. In the control group (n = 9), the animals received sham treatment with distilled water. Serum and liver samples were collected at 6 hours, 3 days and 7 days after treatment. Liver damage was measured using a liver function test and histological analyses. Liver fibrogenesis and hepatic stellate cell (HSC) activation were evaluated using Sirius Red and anti-alpha-smooth muscle actin (α-SMA) immunohistochemical stains.

Results

TACE caused liver injury with greater increases in serum alanine aminotransferase and aspartate aminotransferase levels on day 3 (P<0.05). Histological analyses revealed increased hepatic necrosis in adjacent non-tumorous liver tissue from day 3 compared to the TAE group (Suzuki score of 2.33±1.29 versus 1.13±1.18, P = 0.001). HSC activation and proliferation were significantly increased in the TACE group compared to the control group at 3 and 7 days after treatment (0.074±0.014 vs. 0.010±0.006, and 0.088±0.023 vs. 0.017±0.009, P<0.05). Sirius Red staining demonstrated a statistically significant increase in collagen deposition in the livers in the TACE group 7 days after embolization compared to the control group (0.118±0.012 vs. 0.060±0.017, P = 0.05).

Conclusion

The results of this animal study revealed that TACE induced prominent hepatocellular damage and hepatic fibrogenesis, which compromised liver function and may be responsible for chronic liver decompensation.     

Artemin Stimulates Radio- and Chemo-resistance by Promoting TWIST1-BCL-2-dependent Cancer Stem Cell-like Behavior in Mammary Carcinoma Cells

Artemin (ARTN) has been reported to promote a TWIST1-dependent epithelial to mesenchymal transition of estrogen receptor negative mammary carcinoma (ER-MC) cells associated with metastasis and poor survival outcome. We therefore examined a potential role of ARTN in the promotion of the cancer stem cell (CSC)-like phenotype in mammary carcinoma cells. Acquired resistance of ER-MC cells to either ionizing radiation (IR) or paclitaxel was accompanied by increased ARTN expression. Small interfering RNA (siRNA)-mediated depletion of ARTN in either IR- or paclitaxel-resistant ER-MC cells restored cell sensitivity to IR or paclitaxel. Expression of ARTN was enriched in ER-MC cells grown in mammospheric compared with monolayer culture and was also enriched along with BMI1, TWIST1, and DVL1 in mammospheric and ALDH1+ populations. ARTN promoted mammospheric growth and self-renewal of ER-MC cells and increased the ALDH1+ population, whereas siRNA-mediated depletion of ARTN diminished these CSC-like cell behaviors. Furthermore, increased ARTN expression was significantly correlated with ALDH1 expression in a cohort of ER-MC patients. Forced expression of ARTN also dramatically enhanced tumor initiating capacity of ER-MC cells in xenograft models at low inoculum. ARTN promotion of the CSC-like cell phenotype was mediated by TWIST1 regulation of BCL-2 expression. ARTN also enhanced mammosphere formation and the ALDH1+ population in estrogen receptor-positive mammary carcinoma (ER+MC) cells. Increased expression of ARTN and the functional consequences thereof may be one common adaptive mechanism used by mammary carcinoma cells to promote cell survival and renewal in hostile tumor microenvironments.