针对实时行人检测场景存在遮挡、形态姿势不同的行人目标,YOLOv5模型对于这些目标检测有明显的漏检问题,提出一种像素差异度注意力机制(pixel difference attention,PDA),不同于传统的通道注意力机制用全局均值池化(global average pool...针对实时行人检测场景存在遮挡、形态姿势不同的行人目标,YOLOv5模型对于这些目标检测有明显的漏检问题,提出一种像素差异度注意力机制(pixel difference attention,PDA),不同于传统的通道注意力机制用全局均值池化(global average pooling,GAP)、全局最大值池化(global max pooling,GMP)来概括整张特征图的信息,全局池化将空间压缩成一个值来表征整个通道,造成了空间信息的流失,PDA将空间信息沿高和宽分别压缩,并将其分别与通道信息联系起来做注意力加权操作,同时提出一种新的通道描述指标表征通道信息,增强空间信息与通道信息的交互,使模型更容易关注到综合了空间和通道维度上的特征图的重要信息,在主干网络末端插入PDA后使模型平均精度(mean average precision,mAP)0.5提升了2.4个百分点,mAP0.5:0.95提升了4.4个百分点;针对实时检测场景的部署和检测速度要求模型拥有较少的参数量和计算量,因此提出了新的轻量化特征提取模块AC3代替原YOLOv5模型中的C3模块,该模块使插入PDA后的改进模型在精度仅仅损失0.2个百分点的情况下,参数量(parameters,Param.)减少了20%左右,浮点运算量(giga floating-point operations,GFLOPs)减少了30%左右。实验结果表明,最终的改进模型比YOLOv5s原模型在VOC行人数据集上mAP0.5提升了2.2个百分点,mAP0.5:0.95提升了3.1个百分点,且参数量减少了20%左右,浮点运算量减少了30%左右,在GTX1050上的检测速度(frames per second,FPS)提升了4。展开更多
背景:溶质载体家族1成员5(solute carrier family 1 member 5,SLC1A5)在多种疾病中发挥了潜在作用,但确切作用机制尚不清楚。构建稳定的SLC1A5过表达和敲低细胞模型可为深入研究SLC1A5在疾病中的确切作用机制以及发现潜在治疗靶点提供...背景:溶质载体家族1成员5(solute carrier family 1 member 5,SLC1A5)在多种疾病中发挥了潜在作用,但确切作用机制尚不清楚。构建稳定的SLC1A5过表达和敲低细胞模型可为深入研究SLC1A5在疾病中的确切作用机制以及发现潜在治疗靶点提供有力的实验工具。目的:构建小鼠SLC1A5过表达和敲低的慢病毒载体,以建立稳定转染的RAW264.7细胞株,为深入探讨SLC1A5在炎症中的作用提供实验基础。方法:根据SLC1A5基因序列设计合成引物并使用聚合酶链反应扩增该基因片段。将目的基因定向接入经Age I/Nhe I酶切的载体质粒GV492中构建重组慢病毒质粒,对阳性克隆进一步筛选后测序比对结果;pHelper1.0质粒载体、pHelper2.0质粒载体、目的质粒载体与293T细胞共同培养并转染,获得慢病毒原液进行包装和滴度测定;在此基础上,通过体外培养RAW264.7细胞,确定嘌呤霉素工作质量浓度;不同滴度的慢病毒分别与RAW264.7细胞共同培养,根据荧光强度确定转染效率;用嘌呤霉素挑选出稳定转染细胞,实时荧光定量聚合酶链反应和蛋白免疫印迹方法检测稳定转染细胞株的SLC1A5基因和蛋白表达水平。结果与结论:(1)测序序列与目的序列一致提示重组慢病毒载体构建成功;(2)过表达SLC1A5慢病毒的滴度为1×10~9 TU/mL,敲低SLC1A5慢病毒的滴度为3×10~9 TU/mL;(3)确定RAW264.7细胞嘌呤霉素工作质量浓度为3μg/mL;(4)过表达/敲低SLC1A5慢病毒转染RAW264.7细胞的最佳条件皆为HiTransG P转染增强液且感染复数值等于50;(5)过表达SLC1A5稳转细胞株中SLC1A5基因和蛋白的表达量明显上调,而敲低SLC1A5稳转细胞株中SLC1A5基因和蛋白的表达量显著下调。结果表明,成功构建了小鼠SLC1A5过表达和敲低的慢病毒载体并获得稳定转染的RAW264.7细胞株。展开更多
Rab5 is a GTPase protein that is involved in intracellular membrane trafficking. It functions by binding to various effector proteins and regulating cellular responses, including the formation of transport vesicles an...Rab5 is a GTPase protein that is involved in intracellular membrane trafficking. It functions by binding to various effector proteins and regulating cellular responses, including the formation of transport vesicles and their fusion with the cellular membrane. Rab5 has been reported to play an important role in the development of the zebrafish embryo;however, its role in axonal regeneration in the central nervous system remains unclear. In this study, we established a zebrafish Mauthner cell model of axonal injury using single-cell electroporation and two-photon axotomy techniques. We found that overexpression of Rab5 in single Mauthner cells promoted marked axonal regeneration and increased the number of intra-axonal transport vesicles. In contrast, treatment of zebrafish larvae with the Rab kinase inhibitor CID-1067700markedly inhibited axonal regeneration in Mauthner cells. We also found that Rab5 activated phosphatidylinositol 3-kinase(PI3K) during axonal repair of Mauthner cells and promoted the recovery of zebrafish locomotor function. Additionally, rapamycin, an inhibitor of the mechanistic target of rapamycin downstream of PI3K, markedly hindered axonal regeneration. These findings suggest that Rab5 promotes the axonal regeneration of injured zebrafish Mauthner cells by activating the PI3K signaling pathway.展开更多
文摘针对实时行人检测场景存在遮挡、形态姿势不同的行人目标,YOLOv5模型对于这些目标检测有明显的漏检问题,提出一种像素差异度注意力机制(pixel difference attention,PDA),不同于传统的通道注意力机制用全局均值池化(global average pooling,GAP)、全局最大值池化(global max pooling,GMP)来概括整张特征图的信息,全局池化将空间压缩成一个值来表征整个通道,造成了空间信息的流失,PDA将空间信息沿高和宽分别压缩,并将其分别与通道信息联系起来做注意力加权操作,同时提出一种新的通道描述指标表征通道信息,增强空间信息与通道信息的交互,使模型更容易关注到综合了空间和通道维度上的特征图的重要信息,在主干网络末端插入PDA后使模型平均精度(mean average precision,mAP)0.5提升了2.4个百分点,mAP0.5:0.95提升了4.4个百分点;针对实时检测场景的部署和检测速度要求模型拥有较少的参数量和计算量,因此提出了新的轻量化特征提取模块AC3代替原YOLOv5模型中的C3模块,该模块使插入PDA后的改进模型在精度仅仅损失0.2个百分点的情况下,参数量(parameters,Param.)减少了20%左右,浮点运算量(giga floating-point operations,GFLOPs)减少了30%左右。实验结果表明,最终的改进模型比YOLOv5s原模型在VOC行人数据集上mAP0.5提升了2.2个百分点,mAP0.5:0.95提升了3.1个百分点,且参数量减少了20%左右,浮点运算量减少了30%左右,在GTX1050上的检测速度(frames per second,FPS)提升了4。
基金supported by the Research Funds of the Center for Advanced Interdisciplinary Science and Biomedicine of IHM,No.QYZD20220002the National Natural Science Foundation of China,No.82071357a grant from the Ministry of Science and Technology of China,No.2019YFA0405600 (all to BH)。
文摘Rab5 is a GTPase protein that is involved in intracellular membrane trafficking. It functions by binding to various effector proteins and regulating cellular responses, including the formation of transport vesicles and their fusion with the cellular membrane. Rab5 has been reported to play an important role in the development of the zebrafish embryo;however, its role in axonal regeneration in the central nervous system remains unclear. In this study, we established a zebrafish Mauthner cell model of axonal injury using single-cell electroporation and two-photon axotomy techniques. We found that overexpression of Rab5 in single Mauthner cells promoted marked axonal regeneration and increased the number of intra-axonal transport vesicles. In contrast, treatment of zebrafish larvae with the Rab kinase inhibitor CID-1067700markedly inhibited axonal regeneration in Mauthner cells. We also found that Rab5 activated phosphatidylinositol 3-kinase(PI3K) during axonal repair of Mauthner cells and promoted the recovery of zebrafish locomotor function. Additionally, rapamycin, an inhibitor of the mechanistic target of rapamycin downstream of PI3K, markedly hindered axonal regeneration. These findings suggest that Rab5 promotes the axonal regeneration of injured zebrafish Mauthner cells by activating the PI3K signaling pathway.
