枯草芽孢杆菌拮抗病原菌对细胞微丝的影响

本研究旨在探讨枯草芽孢杆菌拮抗2种病原菌(鼠伤寒沙门氏菌和产肠毒素大肠杆菌)对人结肠癌细胞Caco-2细胞中微丝的影响。将枯草芽孢杆菌及上述2种病原菌加到Caco-2细胞上,用异硫氰酸荧光素-鬼笔环肽标记细胞微丝,在倒置荧光显微镜下观察细胞微丝的变化;利用Western blotting检测细胞微丝调节蛋白Rac1蛋白的表达变化。结果显示正常Caco-2细胞内微丝呈散在的平行排列;加鼠伤寒沙门氏菌后微丝呈小区域聚集;加产肠毒素大肠杆菌后细胞微丝重排聚集在细胞中央放射状分布或分布紊乱;单独加枯草芽孢杆菌后细胞微丝排列方式变化不大;同时分别加2种病原菌和枯草芽孢杆菌后,大部分的细胞内微丝排列整齐,只是在少部分细胞内存在微丝的聚集。产肠毒素大肠杆菌、鼠伤寒沙门氏菌可使细胞微丝调节蛋白Rac1的表达显著上调,而枯草芽孢杆菌则可抑制病原菌对Rac1的影响。本试验证明产肠毒素大肠杆菌、鼠伤寒沙门氏菌可影响细胞微丝的表达和分布;而枯草芽孢杆菌及其培养上清液可抑制病原菌的这种破坏作用。     

Ginsenoside Rh1 attenuates unilateral ureteral obstruction-induced renal interstitial fibrosis in rats

AIM:To observe the effects of ginsenoside Rh1 (G-Rh1) on unilateral ureteral obstruction (UUO)-induced renal interstitial fibrosis and to investigate the underlying mechanisms. METHODS:Male Sprague-Dawley (SD) rats (n=40) were divided into the following 4 groups:UUO-operated group (UUO group), sham-operated group (sham group), UUO-operated plus a low dose (50 mg·kg^-1·d^-1) of G-Rh1 treatment (low G-Rh1 group) and UUO-operated plus a high dose (100 mg·kg^-1·d^-1) of G-Rh1 treatment group (high G-Rh1 group). The G-Rh1 treatment was carried out by gastric gavage from the next day after the UUO operation once a day for 2 weeks (14 d). Immediately after the final dose of G-Rh1, 24 h urine was collected for the urine protein test, and then the rats were euthanized. The blood was collected for the blood urea nitrogen (BUN) and serum creatinine (SCr) assays, and the kidney was removed for pathological and biochemical evaluations. RESULTS:The levels of 24 h urine protein did not show any significant diffe-rence among the groups, while significantly increased levels of BUN and SCr in UUO group were observed (P<0.05), which was prevented by the treatment with G-Rh1 at both doses in a dose-dependent manner. Pathological evaluation showed the renal tissue damage was obvious in UUO group, which was improved by the treatment with G-Rh1 at both doses. Immunohistochemcial analysis exhibited that UUO increased renal interstitial transforming growth factor-β₁ (TGF-β₁) expression, which was also inhibited by the treatment with G-Rh1 at both doses(P<0.05). Significantly increased protein expression of renal interstitial collagen type I, α-smooth muscle actin (α-SMA) and connective tissue growth factor (CTGF) in UUO group was detected, which was suppressed by the treatment with G-Rh1 at both doses. CONCLUSION:G-Rh1 improves UUO-induced renal dysfunction and attenuates interstitial fibrosis, which is mediated via modulation of TGF-β₁-related pro-fibrogenic signaling pathway.     

Effect of plumbagin on levels of Nox4/ROS and α-SMA in human hepatic stellate cells

AIM: To observe the effect of plumbagin on the mRNA and protein expression of nicotinamide adenine dinucleotidephosphate oxidase 4 (Nox4), reactive oxygen species (ROS) level and protein expression of α-smooth muscle actin (α-SMA) in the HSC-LX2 cells stimulated with transforming growth factor β1 (TGF-β1) in vitro. METHODS: HSC-LX2 cells were cultured in vitro and divided into blank group, model group, high-, medium- and low-dose (2, 1.5 and 1 μmol/L) plumbagin groups. After incubated with each drug for 72 h, the mRNA expression of Nox4 was detected by RT-PCR. ROS levels were tested by in situ loading probe method. The protein contents of Nox4 and α-SMA were measured by Western blot. RESULTS: Compared with model group, after treated with plumbagin for 72 h, the mRNA expression of Nox4, ROS level and α-SMA protein were significantly decreased in high-and medium-dose plumbagin groups (P<0.01). CONCLUSION: Plumbagin inhibits the activation of HSC-LX2 cells via decreasing the expression of Nox4, thus decreasing ROS levels.     

Thy-1 Expressing Mesenchymal Cells in Rat Nephrogenesis in Correlation with Cells Immunoreactive for α-Smooth Muscle Actin and Vimentin

Thy-1 expression may influence myofibroblast development. Through the epithelial-mesenchymal transition (EMT), injured renal epithelial cells undergo regression to the metanephric mesenchymal phenotype and then acquire a myofibroblastic nature (expressing α-smooth muscle actin; α-SMA). Because the metanephric blastema differentiates into mesenchymal and renal epithelial cells, we investigated Thy-1 immunoexpression during nephrogenesis in F344 rats in correlation with vimentin and α-SMA expressions. Kidney samples were obtained from fetuses on gestation days 18 and 21, neonates on days 1-18 and adults at 6 weeks of age. Mesangial cells in S-shaped bodies and immature and mature glomeruli continuously expressed both Thy-1 and α-SMA during early nephrogenesis (fetuses and neonates on days 1-9). During early nephrogenesis, loosely-arranged blastemal cell-derived mesenchymal cells in the cortex and medulla also exhibited Thy-1 and α-SMA, although the α-SMA expression was weaker than that of Thy-1. Vimentin expression coincided with that of Thy-1. These findings indicate that the derivation of α-SMA-expressing myofibroblastic cells may be related to mesangial or blastemal cells expressing both Thy-1 and α-SMA. Interestingly, there was a difference in Thy-1 expression between cortical and medullary tubulointerstitial cells from late nephrogenesis (neonates on days 12-18) and those from adults in that the cortical cells reacted faintly or negatively to Thy-1, whereas the medullary cells reacted strongly to Thy-1; additionally, bundle-arranged mesenchymal cells that were only observed in the neonates on days 1-12 reacted strongly to α-SMA, but faintly to Thy-1. Blastemal cell-derived mesenchymal cells seem to alter the immunoexpressions of Thy-1 and α-SMA, depending on the conditions which they develop. Thy-1 immunoexpression would be useful for investigation of reverse embryogenesis, which might occur in fibrotic kidneys.     

小地老虎2种不同类型肌动蛋白基因的克隆与序列分析

本研究以鳞翅目夜蛾科昆虫小地老虎［Agrotis ipsilon（Hüfnagel）］3龄幼虫整个虫体为材料提取总RNA,利用RT-PCR和cDNA末端快速扩增技术（RACE）,分别扩增得到2条肌动蛋白的cDNA序列Aiactin1和Aiactin2。GenBank数据库搜索及序列比对结果表明,克隆的2条肌动蛋白基因应属于2种不同类型的肌动蛋白,Aiactin1为肌肉特异型肌动蛋白,Aiactin2为细胞质特异型肌动蛋白。Aiactin1的cDNA序列含有1 469个碱基,Aiactin2的cDNA序列含有1 408个碱基。2条基因的cDNA序列均包括一个1 131个碱基的开放阅读框,编码一个含376个氨基酸的蛋白。Aiactin1肌动蛋白分子量约为41.77 ku,等电点5.22;Aiactin2肌动蛋白分子量约为41.83 ku,等电点5.47。Prosite软件分析结果表明,Aiactin1和Aiactin2肌动蛋白氨基酸序列中都存在3个肌动蛋白特征片段。2个基因的cDNA序列已经登录GenBank并获得登录号。     

泡桐基因文库的构建及actin基因同源顺序的亚克隆

以λ系列EMBL_3DNA为载体,LE392为宿主菌,建立了木本植物毛泡桐(Paulownia tomentosa (Thunb)Steudel)丛枝病抗病品系C161基因文库,命名为[λE_s/PTS(Bam-Sau)]。其包装效价达到1.1×10~6pfu,重组率为98%。用动物肌动蛋白(actin)基因为探针,从这一基因库中筛选出3.9Kb的泡桐DNA同源顺序进行亚克隆。所得结果表明:动物actin基因在木本植物基因组中存在一定的同源性。     

豌豆肌动蛋白基因在大肠杆菌中的表达

高等植物细胞内含有肌动蛋白，但含量较低，难于提取及进行进一步研究，我们用PCR的方法扩增肌动蛋白cDNA，并克隆到表达质粒pTrpHis，然后转化大肠杆菌DH5α，在IPTG诱导下，肌动蛋白表达在包涵体中，在没有IPTG诱导时，肌动蛋白表达在胞液中。     

植物细胞微丝骨架对真菌侵染的反应

植物细胞骨架对真菌侵染的反应是近10年来才兴起的一个新研究领域，简要介绍了该领域研究的进展情况和发展趋势，包括：植物微丝骨架的结构和功能的简介：真菌侵染时植物细胞骨架（主要是做丝骨架）的变化情况，并对微丝骨架的这种反应在植物抗病性中所起的作用进行了探讨。     

改进YOLOv3的复杂环境下红花丝检测方法

天气变化、光照变化、枝叶遮挡等复杂环境给红花丝的快速、准确检测带来挑战,影响红花采摘机器人的作业效率,该研究基于改进YOLOv3提出了一种目标检测算法（GSC-YOLOv3）。首先GSC-YOLOv3采用轻量级网络幻影结构GhostNet替换主干特征提取网络,并在保证良好检测精度的前提下,最大限度压缩算法参数,提高算法速度,从而使用少量参数生成红花丝有效特征;其次使用空间金字塔池化结构（spatial pyramid pooling,SPP）实现特征增强,弥补提取红花丝特征过程中的信息损失;最后将卷积块注意力模块（convolutional block attention module,CBAM）融入特征金字塔结构,以解决特征融合过程中的干扰问题,提高算法的检测效率和精度。检测结果表明：GSC-YOLOv3算法在测试集下的平均精度均值达到91.89%,比Faster R-CNN、YOLOv3、YOLOv4、YOLOv5、YOLOv6、YOLOv7算法分别高12.76、2.89、6.35、3.96、1.87、0.61个百分点;在GPU下的平均检测速度达到51.1 帧/s,均比其他6种算法高。在复杂场景下的对比试验结果表明,所改进算法具有高检测精度及良好的鲁棒性和实时性,对解决红花采摘机器人在复杂环境下红花丝的精准检测具有参考价值。