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.     

Home gardening in Singapore: A feasibility study on the utilization of the vertical space of retrofitted high-rise public housing apartment buildings to increase urban vegetable self-sufficiency

In land-scarce cities, high-rise apartment buildings may provide vertical spaces for natural-light home gardening along corridors, rooftops, balconies as well as façades. The vertical space can improve not only urban environmental sustainability but also food security. Using an experimental approach, we investigated the food production potential of a high-rise public housing apartment building based on different gardening systems, food crops, and sunlight availability. A gardening prototype system for building corridors was shown to increase the unit area yield of corridor gardening by fivefold compared to a commercial trough planter system. Additionally, this commercial trough planter system was mainly for leafy vegetable production, whereas the gardening prototype system for corridors is also suitable for climbing crops, such as legumes and cucurbits. Nevertheless, because of the limited space along corridors of the apartment building and the relatively low-light levels on average, corridor gardening was estimated to meet only 0.5 % of the demand for vegetables of the residents living in the apartment building. Rooftop gardening with shallow growing medium (depth < 15 cm) was estimated to meet 3 % of demand, and façade gardening 43 %, given the larger space available. Although the vegetable production potential in this study was estimated based on a particular typology of public housing apartment buildings in Singapore, our results showed that vegetable production in public housing apartment buildings is feasible, and home gardening can produce a substantial amount of vegetables for consumption if well deployed. Governments of highly urbanized cities may wish to invest in better home garden designs for high-rise public housing apartment buildings and encourage residents’ participation in home gardening, which would increase high-rise greenery coverage and improve urban food system resilience. Future studies should also investigate the environmental sustainability and food safety aspects of home gardening in highly urbanized cities.     

黄精水煎剂对β-淀粉样蛋白诱导的大鼠 学习记忆能力下降的保护作用研究

本研究旨在探讨黄精水煎剂(PCD)对β-淀粉样蛋白(Aβ)致阿尔茨海默病(AD)模型大鼠的保护作用及可能机制。通过Aβ25-35微量注射至大鼠双侧海马CA1区建立AD模型,同时用PCD干预,研究PCD对AD模型大鼠学习记忆能力、海马组织损伤和CA1区神经元中过度磷酸化tau蛋白(p-tau)表达的影响。结果表明,PCD可明显缩短模型大鼠逃避潜伏期,对模型大鼠学习记忆能力损伤有保护作用;能改善大鼠海马神经元损伤,减少CA1区在thr231位点tau蛋白的过度磷酸化。这些结果提示,PCD对学习记忆能力有显著的保护作用,这可能与其减少海马内p-tau-thr231水平和消除组织氧化逆境有关。     

Identification of Eusynstyelamide B as a Potent Cell Cycle Inhibitor Following the Generation and Screening of an Ascidian-Derived Extract Library Using a Real Time Cell Analyzer

Ascidians are marine invertebrates that have been a source of numerous cytotoxic compounds. Of the first six marine-derived drugs that made anticancer clinical trials, three originated from ascidian specimens. In order to identify new anti-neoplastic compounds, an ascidian extract library (143 samples) was generated and screened in MDA-MB-231 breast cancer cells using a real-time cell analyzer (RTCA). This resulted in 143 time-dependent cell response profiles (TCRP), which are read-outs of changes to the growth rate, morphology, and adhesive characteristics of the cell culture. Twenty-one extracts affected the TCRP of MDA-MB-231 cells and were further investigated regarding toxicity and specificity, as well as their effects on cell morphology and cell cycle. The results of these studies were used to prioritize extracts for bioassay-guided fractionation, which led to the isolation of the previously identified marine natural product, eusynstyelamide B (1). This bis-indole alkaloid was shown to display an IC₅₀ of 5 µM in MDA-MB-231 cells. Moreover, 1 caused a strong cell cycle arrest in G2/M and induced apoptosis after 72 h treatment, making this molecule an attractive candidate for further mechanism of action studies.     

Effects of JAG1 gene silencing on proliferation and apoptosis of human breast cancer MDA-MB-231 cells

AIM:To investigate the effects of Jagged 1 (JAG1) gene silencing on the proliferation and apoptosis of human breast cancer MDA-MB-231 cells. METHODS:The specific recombinant vector pRS-JAG1 was transfected into MDA-MB-231 cells with lipofectamine. The protein expression of JAG1 was observed by Western blotting after transfection. MTT assay was used to detect the effect of JAG1 gene silencing on the growth of the cells. The apoptosis and cell cycle were analyzed by flow cytometry. The protein levels of cyclin D1, p21CIP1/WAF1, p27KIP1, p-Rb, Bcl-2, Bax, Bcl-xL and cleaved caspase-3 were determined by Western blotting. RESULTS:Compared with control group, the expression level of JAG1 was reduced by pRS-JAG1 transfection for 72 h (P<0.05). The growth of MDA-MB-231 cells in shJAG1 group was significantly inhibited (P<0.05). The percentages of G 0/G 1-phase cells and early apoptotic rate were obviously higher in shJAG1 group than those in control group (P<0.05). The shRNA-mediated JAG1 silencing decreased the protein levels of cyclin D1, p-Rb, Bcl-2 and Bax, and increased the protein levels of p21CIP1/WAF1, p27KIP1, Bax and cleaved caspase-3 (P<0.05). CONCLUSION:JAG1 silencing effectively inhibits the proliferation and induces the apoptosis of human breast cancer cells, suggesting that JAG1 might serve as a therapeutic target for triple-negative breast cancer.     

一株韦司梅拟盘多毛孢的鉴定及其发酵液抗乳腺癌活性

【目的】对1株分离自桃树枝的内生真菌N1进行分类鉴定与系统发育分析,探讨其深层发酵液的抗乳腺癌活性,为该菌资源的进一步开发利用提供依据。【方法】通过形态学特征和ITS rDNA序列对野生菌株N1进行分类鉴定;基于ITS1-5.8S-ITS2序列的变异程度对菌株N1遗传多样性进行分析;采用MTT法检测菌株发酵液石油醚、乙酸乙酯、正丁醇和甲醇提取物的抗乳腺癌活性;观察乙酸乙酯提取物不同作用时间下乳腺癌细胞的形态变化;采用LC-MS/MS对乙酸乙酯活性提取物的成分进行分析。【结果】根据菌株形态学和ITS rDNA序列特征,将该野生菌株N1鉴定为韦司梅拟盘多毛孢Pestalotiopsis vismiae;其ITS1-5.8S-ITS2序列总变异率为10.3%,表明韦司梅拟盘多毛孢在自然界中具有丰富的遗传多样性。菌株N1深层发酵液乙酸乙酯提取物对人乳腺癌MDA-MB-231细胞生长的抑制作用最强(P0.05),作用24h的半数抑制浓度(IC50)为131.39μg/mL,作用48h时,细胞呈现凋亡状态。LC-MS/MS数据表征了其中的13个化学成分,确定了6种物质,对其他未能鉴定的7个物质的分子式进行了初步表征。【结论】获得了韦司梅拟盘多毛孢遗传多样性的基础信息,其深层发酵液乙酸乙酯提取物是发挥抗乳腺癌细胞的主要活性物质。     

miR-193b enhances cytotoxicity of doxorubicin by targeting Mcl-1 in breast cancer

AIM: To investigate the effect of microRNA(miR)-193b on doxorubicin therapy in breast cancer in vitro.METHODS: miR-193b level in plasma was detected by real-time PCR in the patients with breast cancer or the healthy controls. MTT assay was performed to measure the inhibitory effect of miR-193b plus doxorubicin on the growth of MDA-MB-231 cells. Bioinformatics, real-time PCR and Western blot were performed to determine whether the expression of Mcl-1 was regulated by miR-193b. Mcl-1 expression vector was constructed, and the role of Mcl-1 vector toward miR-193b plus doxorubicin-induced cytotoxicity in MDA-MB-231 cells was observed by MTT assay.RESULTS: Down-regulation of miR-193b was found in breast cancer patients. The miR-193b plus doxorubicin group showed a higher growth inhibition than cisplation group in MDA-MB-231 cells. The expression of Mcl-1 at both mRNA and protein levels was down-regulated after miR-193b transfection. The growth inhibition of MDA-MB-231 cells treated with miR-193b plus doxorubicin was significantly decreased after the transfection of Mcl-1 expression vector.CONCLUSION: miR-193b sensitizes doxorubicin-induced cytotoxicity by targeting Mcl-1 in breast cancer.     

复合螺旋藻多糖对人乳腺癌231细胞抑制作用的研究

[目的]研究复合螺旋藻多糖对人乳腺癌231细胞的抑制作用。[方法]将螺旋藻多糖（PSP）与银杏叶有效成分（GBE）按一定比例复合,观察复合制剂对人乳腺癌231细胞的抑制作用。[结果]与空白对照组相比,PsP与GBE按1：1、1：2和2：1比例的复合制剂组对人乳腺癌231细胞的抑瘤率,高剂量时分别提高93．32％、57．83％和44．95％,中剂量时分别提高67．52％、59．93％和41．72％,低剂量时分别提高54．22％、48．56％和40．26％。单一成分的螺旋藻多糖组对人乳腺癌231细胞有较好的抑制作用,但随着剂量地降低其药效也随之降低。[结论]复合螺旋藻多糖对人乳腺癌231细胞有较高地抑制作用,其中,以高剂量组1：1复合制剂抑制效果最好（93．32％）,表明复合螺旋藻多糖在该剂量配比时协同增效作用达到最大。     

Surface mould growth on wooden claddings – effects of transient wetting,relative humidity,temperature and material properties

ABSTRACT

Effects of climatic factors and material properties on the development of surface mould growth on wooden claddings were investigated in a laboratory experiment. Specimens of aspen (Populus tremula), Siberian larch (Larix Sibirica), American white oak (Querqus alba), Scots pine (Pinus sylvestris), Norway spruce (Picea abies) and thermally modified pine were incubated in eight climatic chambers at specified wetting periods (2 or 4?h per day), relative humidity (58–86%) and temperature conditions (10–27°C). Surface mould growth was assessed weekly for 13 weeks, and the results were evaluated statistically using Generalized Estimating Equations logistic regression models. All tested climatic factors had significant effects on the mould growth, and there were significant differences between the materials. The ranking of the materials varied with temperature and over time. Aspen, pine sapwood and oak were overall most susceptible to mould growth, and thermally modified pine least susceptible. There were significant differences between sapwood and heartwood for pine and spruce. The effect of density was tested on the spruce heartwood material, but was not found to be significant. The results can be used to further develop prediction models for mould growth on wooden claddings.     

A shot in the dark for conservation: Evidence of illegal commerce in endemic and threatened species of elasmobranch at a public fish market in southern Brazil

1. The illegal sale of endangered elasmobranchs has been recorded in a number of different Brazilian states, where sharks and rays are being marketed primarily as ‘cação’ or ‘viola’. Brazil is ranked among the top 10 nations worldwide that harvest most sharks, causing an immeasurable impact on the local elasmobranch populations.
2. The present study applied the DNA barcoding technique, based on the cytochrome C oxidase subunit I gene, for the molecular identification of the elasmobranch species sold as processed products under the generic names of ‘cação’, ‘mangona’, ‘azul’, ‘cambeva’, ‘fiuso’ and ‘lombo preto’, in the fish market of the city of Florianópolis, capital of the state of Santa Catarina, Brazil.
3. Nine elasmobranch species were identified in the 56 samples analysed, including six sharks and three rays, representing six families, the Carcharhinidae, Sphyrnidae, Squatinidae, Arhynchobatidae, Myliobatidae and Gymnuridae. Prionace glauca, identified in more than half (56%) of the samples analysed, is listed as Near Threatened by the International Union for the Conservation of Nature.
4. Six species identified in the present study, Sphyrna zygaena, Sphyrna lewini, Squatina guggenheim, Carcharhinus signatus, Gymnura altavela and Rioraja agassizii, are under some level of risk of extinction, while two others (Rhizoprionodon lalandii and Myliobatis goodei) are listed as Data Deficient.
5. Our results indicate that the commercial exploitation of endemic sharks and rays at risk of extinction is commonplace in southern Brazil. This reinforces the need for more systematic monitoring of the trade in fishery products and more effective application of the environmental legislation and conservation programmes.