PTPN6通过抑制SP1/p38 MAPK信号通路促进前列腺癌细胞的化疗敏感性

目的:研究PTPN6对前列腺癌细胞PC3的作用及其作用机制。方法:RT-PCR和Western blot实验检测前列腺癌组织和细胞以及癌旁组织和人前列腺上皮细胞中PTPN6的表达量；CCK-8和EDU染色实验检测PTPN6对前列腺癌细胞PC3增殖的影响；Western blot实验检测耐药相关蛋白P-gp和MRP-1的蛋白表达水平。结果:RT-PCR和Western blot结果显示，PTPN6在前列腺癌组织和细胞中的表达量显著低于癌旁组织和人前列腺上皮细胞中的表达量；过表达PTPN6显著抑制前列腺癌PC3细胞的增殖，并降低PC3细胞的耐药性；进一步的研究结果表明PTPN6可通过抑制SP1，并抑制p38 MAPK通路抑制PC3细胞的增殖和耐药。结论:PTPN6能够抑制前列腺癌细胞PC3的增殖和耐药，提高其化疗敏感性，作用机制是通过调控SP1/p38 MAPK信号通路来实现的，这一结果能够为临床上前列腺癌的诊断和治疗提供分子基础。     

B cell and B cell-related pathways for novel cancer treatments

B cells are recognized as the main effector cells of humoral immunity which suppress tumor progression by secreting immunoglobulins, promoting T cell response, and killing cancer cells directly. Given these properties, their anti-tumor immune response in the tumor micro-environment (TME) is of great interest. Although T cell-related immune responses have become a therapeutic target with the introduction of immune checkpoint inhibitors, not all patients benefit from these treatments. B cell and B cell-related pathways (CCL19, −21/CCR7 axis and CXCL13/CXCR5 axis) play key roles in activating immune response through humoral immunity and local immune activation via tertiary lymphoid structure (TLS) formation. However they have some protumorigenic works in the TME. Thus, a better understanding of B cell and B cell-related pathways is necessary to develop effective cancer control. In this review, we summarize recent evidences regarding the roles of B cell and B cell-related pathways in the TME and immune response and discuss their potential roles for novel cancer treatment strategies.     

赤芝FPS基因酵母单杂交文库构建及其上游转录因子的筛选

目的筛选赤芝三萜合成途径中法尼基焦磷酸合酶(farnesyl diphosphate synthase,FPS)基因的上游调控转录因子。方法首先克隆了FPS基因启动子,并连接至pAbAi质粒构建诱饵载体pAbAi-FPS。将pAbAi-FPS转化Y1H酵母感受态细胞构建诱饵菌。采用SMART技术构建赤芝酵母单杂交cDNA文库,再将纯化的双链cDNA、pGADT7-Rec共转化诱饵菌株,筛选FPS上游的转录调控因子。结果构建了含pAbAi-FPS的诱饵载体并筛选出诱饵菌株,构建了cDNA文库并转化诱饵菌株,筛选出阳性克隆37个,得到作用FPS基因上游的转录调控因子18个,包括转录因子3个、核糖体蛋白5个及其他家族蛋白10个。结论筛选出转录因子GlSNF2、GlMHR和GlZn2Cys6为调控FPS表达的候选基因,为深入研究FPS基因的表达调控机制奠定了研究基础。     

SDF-1α/CXCR4 信号通路在轴向应力刺激促进骨再生中的作用研究

目的通过轴向应力刺激促进骨再生，观察基质细胞衍生因子 1α/趋化因子 CXC 亚族受体 4（stromal cell-derived factor 1α/cysteine X cysteine receptor 4，SDF-1α/CXCR4）信号通路变化，探讨轴向应力刺激促进骨再生的机制。方法取 72 只雄性新西兰大白兔，于右后肢胫骨近端内侧制备直径 8 mm 圆形皮质骨缺损并脱蛋白松质骨支架修复模型，随机分为 3 组（n=24）。A 组腹腔注射 PBS，B 组术肢给予应力刺激治疗+腹腔注射 PBS，C 组术肢给予应力刺激治疗+腹腔注射 CXCR4 拮抗剂（AMD3100）。术后 2、4、8、12 周，摄 X 线片并采用 Lane-Sandhu X 线评分标准评价骨愈合情况，取标本行 HE 染色观察新生骨组织及支架降解，免疫组织化学染色观察 VEGF、CXCR4 表达水平；4、8 周取标本 Western blot 检测 SDF-1α 及 CXCR4 蛋白表达水平；12 周行 Micro-CT 检查，计算新生骨体积及新生骨密度。 结果X 线片检查示，除术后 2 周各组骨缺损区及支架无明显变化外，4、8 及 12 周时 B 组骨愈合评分均高于 A、C 组（P<0.05）。12 周时 Micro-CT 扫描可见 B 组骨缺损修复、髓腔再通，新生骨体积及骨密度均高于 A、C 组（P<0.05）。HE 染色显示，术后 4 周开始 B 组骨再生及支架降解均明显快于 A、C 组。免疫组织化学染色示，各组 VEGF 及 CXCR4 阳性表达均在 4 周达峰值；各时间点 B 组 VEGF 及 CXCR4 表达量均显著高于 A、C 组（P<0.05）。Western blot 检测显示，4、8 周时 B 组 SDF-1α 与 CXCR4 表达量均显著高于 A、C 组（P<0.05）。 结论轴向应力刺激促进骨再生可能与其促进骨缺损区组织高表达 SDF-1α，激活与其下游调控 BMSCs 募集的 CXCR4 信号有关。     

Gender equity: how do the forensic sciences fare?

ABSTRACT

Females are underrepresented in science, technology, engineering and mathematics (STEM) at all levels of society. Fewer females are completing STEM school subjects, graduating with STEM degrees, being employed as STEM professionals, and holding senior leadership and academic positions in STEM. However, unlike almost every other STEM discipline, the overall ratio of females is higher in many forensic science disciplines. For our sector, rather than having difficulty in attracting females, the bigger issue is how we retain and promote female talent. This complex issue is exacerbated by: gender pay gaps; family role expectations; lack of visible role models or mentors; discrimination and harassment; and bias during recruitment and promotion practices. We discuss barriers relevant for women in the forensic industry and offer potential solutions. These include flexible work arrangements, sponsorship programmes, and fostering and practising an inclusive workplace culture. Gender equity programmes and exemplar STEM organizations focused on a commitment to gender parity will be explored. Harnessing untapped female talent is as much a social justice issue as employing best practices for improving the quality, diversity and output of our forensic science workforce, and research and innovation strategies.     

Impact of various culture conditions on ex vivo expansion of polyclonal T cells for adoptive immunotherapy

Currently, adoptive immunotherapy is considered as one of the leading treatments in cancer. Successful adoptive immunotherapy depends on producing large numbers of desired T cells ex vivo for infusion. This requires an effective protocol for maximum functional T‐cell expansion while keeping the time and costs to a minimum. Current T‐cell expansion protocols are diverse in their methodology, and a universal protocol of expansion is wanting. Also, new findings regarding T‐cell biology, signaling, and activation have reshaped the strategies of T‐cell propagation over the years, introducing new ways to expand T cells. Here, we reviewed different conditions for blood‐derived polyclonal T‐cell expansion so as to elucidate the influential factors of T‐cell expansion and their efficacy.     

Predictions of genotoxic potential,mode of action,molecular targets,and potency via a tiered multiflow® assay data analysis strategy

The in vitro MultiFlow® DNA Damage Assay multiplexes γH2AX, p53, phospho-histone H3, and polyploidization biomarkers into a single flow cytometric analysis. The current report describes a tiered sequential data analysis strategy based on data generated from exposure of human TK6 cells to a previously described 85 chemical training set and a new pharmaceutical-centric test set (n = 40). In each case, exposure was continuous over a range of closely spaced concentrations, and cell aliquots were removed for analysis following 4 and 24 hr of treatment. The first data analysis step focused on chemicals' genotoxic potential, and for this purpose, we evaluated the performance of a machine learning (ML) ensemble, a rubric that considered fold increases in biomarkers against global evaluation factors (GEFs), and a hybrid strategy that considered ML and GEFs. This first tier further used ML output and/or GEFs to classify genotoxic activity as clastogenic and/or aneugenic. Test set results demonstrated the generalizability of the first tier, with particularly good performance from the ML ensemble: 35/40 (88%) concordance with a priori genotoxicity expectations and 21/24 (88%) agreement with expected mode of action (MoA). A second tier applied unsupervised hierarchical clustering to the biomarker response data, and these analyses were found to group certain chemicals, especially aneugens, according to their molecular targets. Finally, a third tier utilized benchmark dose analyses and MultiFlow biomarker responses to rank genotoxic potency. The relevance of these rankings is supported by the strong agreement found between benchmark dose values derived from MultiFlow biomarkers compared to those generated from parallel in vitro micronucleus analyses. Collectively, the results suggest that a tiered MultiFlow data analysis pipeline is capable of rapidly and effectively identifying genotoxic hazards while providing additional information that is useful for modern risk assessments—MoA, molecular targets, and potency. Environ. Mol. Mutagen. 60:513–533, 2019. © 2019 Wiley Periodicals, Inc.