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1.
病毒侵染直接影响草莓的生长发育及果实品质,快速检测及鉴定病毒是病毒防治的前提。本研究以‘丰香’草莓和‘哈尼’草莓为试材,利用小RNA(sRNA)测序结合RT-PCR技术对2种栽培草莓品种中存在的病毒进行了研究。对sRNA测序结果进行组装和注释,结果表明:在混合样品中共有242个contigs比对到5种草莓病毒,分别为草莓白化病毒(strawberry pallidosis associated virus,SPaV)、草莓镶脉病毒(strawberry vein banding virus,SVBV)、草莓斑驳病毒(strawberry mottle virus,SMoV)、草莓毛形病毒3(strawberry crinivirus 3,SCrV 3)和草莓毛形病毒4(strawberry crinivirus 4,SCrV 4)。利用RT-PCR技术对sRNA测序结果进行验证,结果表明:SMoV、SVBV和SCrV 3在‘丰香’草莓和‘哈尼’草莓中均检测到,而SPaV和SCrV 4只在‘哈尼’草莓样品中检测到。本研究利用sRNA测序技术鉴定了2个草莓品种中存在的病毒,首次在我国生产的同一个草莓品种中检测到5种病毒,为草莓病毒的检测和防控提供了新的思路。  相似文献   
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AIM: To investigate the effect of shikonin on reversing hepatocyte growth factor(HGF)-induced resistance to gefitinib in lung cancer HCC827 cells, and to explore its possible mechanisms.METHODS: The gefitinib-resistant HCC827 cells induced by HGF were treated with shikonin and gefitinibthe alone or in combination. The inhibition rates of cell viability were determined by MTT assay. The invasive ability of HCC827 cells with HGF-induced resistance to gefitinib was determined by Transwell assay. The protein levels of epithelial-mesenchymal transition (EMT) and related signaling pathway in the HCC827 cells were detected by Western blot.RESULTS: The results of MTT assay showed that the cell activity of HCC827 cells was significantly inhibited by shikonin in a dose dependent manner. The IC50 of shikonin in HCC827 cells was 3.06 μmol/L. And the IC50 of gefitinib in HCC827 cells was 0.51 μmol/L. Under the condition of combined treatment with shikonin and gefitinib in the presence of HGF (20 μg/L), the IC50 of gefitinib was 7.36 μmol/L, significantly lower than that treated with gefitinib alone (P<0.01), so did the result of the cell migration (P<0.01). HGF induced EMT, while shikonin reversed this effect. The protein expression level of p-AKT was significantly up-regulated by HGF, while markedly down-regulated treatment with shikonin and gefitinib compared with gefitinib alone (P<0.01).CONCLUSION: Shikonin reverses HGF-induced resistance to gefitinib in lung cancer HCC827 cells, and the mechanism may be likely related to the preventon of EMT and the inhibition of HGF-induced activation of p-AKT signaling pathway.  相似文献   
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Urban Norway rat (Rattus norvegicus) populations can carry the bacteria methicillin‐resistant Staphylococcus aureus (MRSA). There are numerous knowledge gaps in the epidemiology of MRSA in these populations that limit understanding of its ecology in urban environments. For example, fecal shedding of MRSA, which may increase environmental contamination, has been reported in other species; however, it is unknown whether Norway rats carry the bacteria rectally. Furthermore, while intermittent MRSA shedding has been shown in other species and may dictate when the risk of transmission is highest, duration of carriage has not been examined for Norway rats. Previous work has shown that lethal animal‐control methods may increase the level of pathogens within reservoir populations, possibly by disrupting ecological patterns. However, the impact of rodent‐control on potentially environmentally acquired pathogens like MRSA has not been tested. Using capture‐mark‐recapture methods in an inner‐city neighborhood in Vancouver, Canada, we show that rats intermittently carry MRSA both in the rectum and oropharynx. By assessing the prevalence of MRSA before and after enacting a pest‐control intervention, we report that kill‐trapping had no impact on the prevalence of carriage of this environmentally‐acquired agent.  相似文献   
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AIM: To investigate the effect of naringin (NRG) on cisplatin (DDP) resistance in human lung cancer A549/DDP cells and its possible mechanism. METHODS: A549/DDP cells were cultured in vitro and treated with NRG and/or DDP at different concentrations for 24 h, and then the cell viability were measured by CCK-8 assay. The combination index (CI) of NRG and DDP were analyzed by Chou-Talalay method. The apoptosis rate was analyzed by flow cytometry. Western blot was performed to detect the protein levels of P-glycoprotein (P-gp), multidrug resistance-associated protein 1 (MRP1), p-Akt, CXC chemokine receptor 4 (CXCR4), cleaved caspase-3, Bcl-2 and Bax.RESULTS: The protein levels of P-gp, MRP1, p-Akt and CXCR4 in the A549/DDP cells were higher than those in the A549 cells (P<0.05). The cell viability was remarkably reduced in a dose-dependent manner when A549/DDP cells were exposed to NRG and/or DDP (P<0.05), and the IC50 values of NRG and DDP were 36.92 μmol/L and 129.77 μmol/L, respectively. When the inhibition rate exceeded 15%, NRG in combination with DDP produced a synergistic effect (CI<1). Combination treatment with NRG and DDP significantly induced apoptosis (P<0.05), up-regulated the protein levels of cleaved caspase-3 and Bax, and down-regulated the protein level of Bcl-2 (P<0.05). Meanwhile, NRG remarkably down-regulated the protein levels of P-gp, MRP1, p-Akt and CXCR4 in a dose-dependent manner (P<0.05). CONCLUSION: NRG may enhance the sensibility of A549/DDP cells to DDP most likely via up-regulating the protein level of Bax and down-regulating the protein levels of Bcl-2, P-gp, MRP1, p-Akt and CXCR4.  相似文献   
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AIM: To investigate the effect of Linc00152 on the viability, apoptosis and radiosensitivity of cervical cancer cells. METHODS: RT-qPCR was used to detect the expression levels of Linc00152 and microRNA-376c-3p(miR-376c-3p) in human cervical cancer HeLa cells and SiHa cells, and normal cervical Ect1/E6E7 cells. The cervical cancer HeLa cells with low Linc00152 expression or miR-376c-3p over-expression were established. MTT assay, flow cytometry, colony formation assay and Western blot were used to determine the cell viability, apoptosis, radiosensitivity and related protein expression. The dual-luciferase reporter assay was used to verify the regulatory relationship between Linc00152 and miR-376c-3p in the HeLa cells. RESULTS: Compared with the Ect1/E6E7 cells, Linc00152 was up-regulated in the HeLa cells and SiHa cells, and miR-376c-3p was down-regulated (P < 0.05). Low expression of Linc00152 or over-expression of miR-376c-3p inhibited the viability of HeLa cells, induced apoptosis, enhanced the radiosensitivity, inhibited the protein expression of cyclin D and Bcl-2, and promoted the protein expression of P21 and Bax (P < 0.05). Linc00152 negatively regulated miR-376c-3p expression in the HeLa cells, and inhibition of miR-376c-3p expression reversed the effect of low expression of Linc00152 on HeLa cell viability, apoptosis and radiosensitivity. CONCLUSION: Linc00152 is highly expressed in the cervical cancer cells. Linc00152 affects the viability, apoptosis and radiosensitivity of HeLa cells by targeting miR-376c-3p, which is a potential diagnosis and treatment target for cervical cancer.  相似文献   
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AIM To investigate the effect of exosomes secreted by mouse melanoma cells on the expression of Ras-related C3 botulinum toxin substrate 1 (Rac1) protein in fibroblasts. METHODS Ultracentrifugation was adopted to separete exosomes secreted by mouse melanoma B16-F10 cells. The morphological structure of exosomes was observed by negative-staining electron microscopy. The size distribution of exosomes was determined by nanoparticle tracking analysis (NTA). The exosomal markers, tumor susceptibility gene 101 (Tsg101) and tyrosinase-related protein 2 (Tyrp2), were identified by Western blot. Laser confocal microscopy was used to observe the process that mouse embryonic fibroblasts (MEF) took in exosomes during co-culture. Immunocytochemical staining and Western blot were used to detect the expression of Rac1 protein in MEF. RESULTS B16-F10 cell exosomes showed a typical tea tray-like structure, with a size range of 141~255 nm, and expressed protein markers Tsg101 and Tyrp2. The results of laser confocal microscopy showed that compared with co-culture at 0 h, a small number of exosomes appeared in the MEF at 12 h, and a large number of exosomes accumulated in the MEF after co-cultured for 24 and 36 h. Western blot analysis showed that compared with co-culture at 0 h, the expression of Rac1 protein in the MEF was significantly increased at 24 h and 36 h of co-culture (P<0.01). The results of immunocytochemical staining showed that compared with co-culture at 0 h, the positive expression level of Rac1 in the MEF cells was significantly increased at 12 h, 24 h and 36 h of co-culture (P<0.05 or P<0.01). CONCLUSION Intake of exosomes secreted by mouse melanoma cells promotes the expression of Rac1 protein in fibroblasts.  相似文献   
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为了探索研究茶氨酸衍生物茶溴香酰胺制备的脂质体(TBrC-L)对人非小细胞肺癌A549细胞生长和迁移的抑制作用与其分子机制,采用薄膜分散法制备了TBrC-L,通过MTT法检测不同浓度的TBrC-L对人肺癌A549细胞生长的抑制作用;使用流式细胞术检测TBrC-L对人肺癌A549细胞凋亡的诱导作用;利用Transwell chamber法观察TBrC-L对人肺癌A549细胞迁移作用的影响;应用蛋白质印迹法检测人肺癌A549细胞中与凋亡和生长密切相关蛋白的表达和药物可能的作用靶点。实验结果显示,TBrC-L对人肺癌A549细胞生长和迁移有显著的抑制作用,促进肿瘤细胞凋亡、抑制肿瘤细胞生长和迁移可能是其抗人肺癌作用的重要机制,其作用的分子机制涉及到抑制EGFR、VEGFR1、VEGFR~2和Met受体介导的Akt和NF-κB信号传导通路,本研究结果提示,TBrC-L具有应用于临床治疗和(或)辅助治疗人肺癌的潜力。  相似文献   
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The prevalence of cancer in animals has increased significantly over the years. Mammary tumours are the most common neoplasia in dogs, in which around 50% are presented in the malignant form. Hence, the development and characterization of in vitro models for the study of canine tumours are important for the improvement of cancer diagnosis and treatment. Thus, the aim of this study was to characterize cell lines derived from canine mammary gland neoplasias which could be further used for basic and applied oncology research. Samples of canine mammary carcinomas were taken for cell culture and 2 cell lines were established and characterized in terms of cell morphology, tumourigenicity and global gene expression. Both cell lines presented spindle‐shape morphology and shown common malignant features as in vitro invasion potential and expression of epithelial and mesenchymal proteins. Also, we found gene expression patterns between the 2 cell cultures in comparison to the normal mammary gland tissue. Cells from M25 culture showed a higher invasion and in vivo tumourigenic potential, associated to the overexpression of genes involved in focal adhesion and extracellular matrix communication, such as FN1, ITGA8 and THBS2. The phenotypic characterization of these cells along with their global gene expression profile potentially determine new therapeutic targets for mammary tumours.  相似文献   
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