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目的构建稳定共表达荧光素酶（Luc）和增强型绿色荧光蛋白（EGFP）的人源卵巢癌细胞SKOV3（SK-Luc-EGFP）并探究其在卵巢癌腹腔转移的体内外研究中的应用。方法利用重叠PCR扩增得到的Luc-T2A-EGFP基因片段，构建重组质粒pCDH-Luc-T2A-EGFP-Puro；采用三质粒慢病毒包装系统转染HEK 293T细胞，收集病毒液感染SKOV3细胞；通过嘌呤霉素筛选，流式细胞术检测，筛选高效表达EGFP的细胞（SK-Luc-EGFP），并通过体外生物发光实验验证Luc的表达。对SK-Luc-EGFP细胞进行如下应用的探究：利用流式细胞术和激光共聚焦区分SK-Luc-EGFP细胞与腹水微环境中的非肿瘤细胞，利用荧光显微镜观察SK-Luc-EGFP与间皮细胞的黏附，利用小动物活体成像仪观察SK-Luc-EGFP细胞的大网膜黏附以及体内成瘤的过程。结果成功构建慢病毒载体pCDH-Luc-T2A-EGFP-Puro；感染并筛选得到SK-Luc-EGFP单克隆细胞株。通过荧光显微镜和流式细胞术均能检测到EGFP的表达，细胞纯度达100%；通过流式细胞术和激光共聚焦成像可直观辨别SK-Luc-EGFP细胞与腹水微环境细胞。体外生物发光试验成功验证Luc的表达，且细胞数和生物发光信号的线性相关系数为0.997 9。细胞黏附试验中通过荧光成像直接观察到SK-Luc-EGFP细胞对间皮细胞的黏附；经腹腔注射SK-Luc-EGFP细胞观察到细胞在裸鼠大网膜上的黏附；在SK-Luc-EGFP细胞建立的卵巢癌腹腔转移模型中，利用小动物活体成像仪实现对腹腔肿瘤的活体监测。结论SK-Luc-EGFP细胞株在卵巢癌腹腔转移的体内外研究中具有多重应用潜力。

Copyright of Journal of Petrochemical Universities / Shiyou Huagong Gaodeng Xuexiao Xuebao is the property of Journal Editorial Department Of Liaoning Shihua University and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

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The amount and timing of biochar application in scientific settings are currently the focal points of research aimed at enhancing and maintaining soil health. In this study, two groups of corn pot experiments（groups A and C）were designed. Group A was treated with a one-time application of biochar for 7 years, and group C was treated with a one-time application of biochar for 1 year. The application rates of each group were 0, 15.75, 31.50, 63.00 t·hm-2, and 126.00 t·hm-2, respectively. The effects of different amounts and timings of biochar application on the structural characteristics of nematode communities in sandy soil were studied. The results showed that the application of biochar could increase soil pH and nutrient content after 1 year. The effect of biomass charcoal on soil nutrient would decrease over time with aging. The dominant genus of soil nematodes was Acrobeloides, and bacterivores and predators / omnivores were the main trophic groups. Soil organic matter decomposition was dominated by bacterial decomposition. The increase in biochar application amount changed the soil nematode community structure, trophic group distribution, and nematode abundance after one year of biochar application. The increase in the amount of biochar applied substantially inhibited the number of soil nematodes. However, the number of soil nematodes recovered after 7 years of biochar application. With increasing years after application, biochar application led to a decrease in the abundance of soil bacterivores and an increase in the abundance of predators/omnivores and plant parasitic nematodes; however, it exerted little effect on the nematode diversity and richness indexes. A one-time application of 126.00 t·hm-2 biochar substantially reduced the number of soil nematodes after 7 years, whereas the application of 15.75 t·hm-2 biochar exerted a positive effect on the number and community structure of soil nematodes. [ABSTRACT FROM AUTHOR]

Chinese Expert Consensus 2023 on Endovascular Treatment for Acute Large Vessel Occlusion with Mild Stroke is a summary and analysis of the new progress and new evidence obtained from the recent studies on endovascular treatment for acute large vessel occlusion with mild stroke by the experts organized by the Chinese Interventional Neuroradiology Society of Chinese Stroke Association. This expert consensus covers the definition of mild stroke, and intravenous thrombolysis and endovascular treatment for acute large vessel occlusion that begins with mild stroke. The aim is to provide guidance for professionals and social workers engaged in stroke prevention and treatment, especially the endovascular treatment, care and rehabilitation of acute ischemic stroke, as well as relevant governmental agencies, healthcare administrators, pharmaceutical companies, healthcare demand parties, and other stakeholders. [ABSTRACT FROM AUTHOR]

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The study on the equity of urban green space layout based on the quantification of ecosystem services is oriented to the improvement of human well-being, which is helpful to promote the equity of ecological environment and the sustainable development of cities. Based on the analysis of the urban green space distribution and ecosystem service value of green space of the blocks in Yuexiu District,Guangzhou in 2020, this study used the location entropy method to analyze the fairness of green space ecosystem services available per capita in each block. The results showed that: (1) The population and urban green space in Yuexiu District were significantly spatially heterogeneous. (2) The comprehensive ecosystem service value of green space varied greatly among the blocks in Yuexiu District, with a high spatial distribution in the north and a low spatial distribution in the south. (3) There were some unfair spatial distribution of green space in the district. The two blocks, Dengfeng and Hongqiao, had extremely high location entropy. The seven blocks of Renmin, Guangta, Zhuguang, Dadong, Datang, Meihua Village and Kuangquan had low or extremely low location entropy. It was proposed to optimize the green space layout of the study area by strictly protecting important ecological green space, inserting green space in subtleties, increasing urban public green space by using urban wasteland, adding riverside greenways with sports and leisure functions, increasing urban green space under the background of old city reconstruction, and increasing the urban vertical green area. The paper results can provide some reference for the study of the fairness of green space layout in the same type of high population density cities, which can help to improve the level of residents' health and well-being. [ABSTRACT FROM AUTHOR]

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Objective To investigate the effect of the SMAC gene on paclitaxel sensitivity and cellular activity in lung adenocarcinoma cells based on the caspase-3/Bcl-2/Bax signaling pathway. Methods A paclitaxel-resistant cell line A549/Taxol was established for lung adenocarcinoma, and the cells were divided into four following groups: pcDNA-NC (transfected with pcDNA-NC blank vector), pcDNA-SMAC (transfected with pcDNA-SMAC vector), siRNA-NC (transfected with siRNA-NC empty virus vector), and siRNA-SMAC groups (transfected with siRNA-SMAC lentiviral vector). The SMAC mRNA expression in cells was detected by qRT-PCR; cell sensitivity was detected by MTT; cell proliferation ability was detected by cloning assay; cell invasion ability was detected by Transwell; apoptosis ability was detected by flow cytometry assay; and caspase-3, Bcl-2 and Bax protein expression in cells were detected by Western blot analysis. Results The SMAC mRNA expression was significantly lower in A549 cells compared with BEAS-2B cells (P<0.05). The SMAC mRNA expression was significantly higher in the pcDNA-SMAC group than that in the pcDNA-NC group cells (P<0.05). The SMAC mRNA expression was significantly lower in the cells of the siRNA-SMAC group (P<0.05) than that in the siRNA-NC group. The SMAC mRNA expression was significantly lower in the cells of the siRNA-SMAC group (P<0.05) than in the siRNA-NC group. Compared with the pcDNA-NC group, the cell IC50, cell clone number, cell invasion ability, and Bcl-2 protein and Bcl-2/Bax ratio were significantly lower in the pcDNA-SMAC group, the cell resistance index reversal was 2.51-fold, and the apoptosis ability and caspase-3, as well as Bax protein expression, were significantly higher (P<0.05). Compared with the siRNA-NC group, cell IC50, cell clone number, cell invasion ability, and Bcl-2 protein and Bcl-2/Bax ratio were significantly higher in the siRNA-SMAC group, and apoptosis ability and caspase-3 and Bax protein expression were significantly lower (P<0.05). Conclusion High expression of SMAC increases paclitaxel sensitivity, inhibits cell growth and invasion, promotes apoptosis in lung adenocarcinoma cells, and has a regulatory effect on the caspase-3/Bcl-2/Bax signaling pathway. [ABSTRACT FROM AUTHOR]

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