Universal two-dimensional labelled probe-mediated melting curve analysis based on multiplex PCR for rapid typing of Plasmodium in a single closed tube

Currently, malaria is still one of the major public health problems commonly caused by the four Plasmodium species. The similar symptoms of malaria and the COVID-19 epidemic of fever or fatigue lead to frequent misdiagnosis. The disadvantages of existing detection methods, such as time-consuming, costly, complicated operation, need for experienced technicians, and indistinguishable typing, lead to difficulties in meeting the clinical requirements of rapid, easy, and accurate typing of common Plasmodium species. In this study, we developed and optimized a universal two-dimensional labelled probe-mediated melting curve analysis (UP-MCA) assay based on multiplex and asymmetric PCR for rapid and accurate typing of five Plasmodium species, including novel human Plasmodium, Plasmodium knowlesi (Pk), in a single closed tube following genome extraction. The assay showed a limit of detection (LOD) of 10 copies per reaction and could accurately distinguish Plasmodium species from intra-plasmodium and other pathogens. Additionally, we proposed and validated different methods of fluorescence quenching and tag design for probes that are suitable for UP-MCA assays. Moreover, the clinical performance of the Plasmodium UP-MCA assay using a base-quenched universal probe was evaluated using 226 samples and showed a sensitivity of 100% (164/164) and specificity of 100% (62/62) at a 99% confidence interval, with the microscopy method as the gold standard. In summary, the UP-MCA assay showed excellent sensitivity, specificity, and accuracy for genotyping Plasmodium species spp. Additionally, it facilitates convenient and rapid Plasmodium detection in routine clinical practice and has great potential for clinical translation.     

伴刀豆球蛋白A和层粘连蛋白与膜受体结合下膜分子运动及微丝组装变化的比较

研究伴刀豆球蛋白A和层粘连蛋白分别与小鼠腹腔巨噬细胞膜受体结合下引起细胞膜分子运动的变化和对微丝组装的影响.结果表明,伴刀豆球蛋白A和层粘连蛋白作用下均导致膜表面蛋白分子的侧向扩散速率减慢,膜脂流动性降低,加快膜内微丝组装并使微丝含量增加.两配体作用下引起细胞上述反应有相似性.     

基于免疫纳米颗粒强化的压电免疫传感器检测大肠杆菌O157︰H7

摘要：【目的】结合纳米技术建立检测大肠杆菌（Escherichia coli）O157︰H7高灵敏检测技术。【方法】采用化学共沉淀法制备出核心粒径约为10 nm的免疫纳米磁颗粒,柠檬酸钠还原法制备粒径约为20 nm的免疫胶体金。压电免疫传感器通过金黄色葡萄球菌蛋白A（Protein A from Staphylococcus aureus SPA）法将抗体固定于石英晶振上,两种免疫纳米颗粒借助不同的抗体连接于传感器上对检测频率信号进行放大。【结果】SPA在石英晶振上的最佳固定浓度和时间为1.2 mg/mL和40 min,抗体的最佳固定浓度和时间为1.0 mg/mL和60 min。压电免疫传感器通过两种免疫纳米颗粒的放大作用,使其对大肠杆菌O157︰H7的检测限从104 cfu/mL提高到101 cfu/mL。【结论】免疫纳米颗粒强化对压电免疫传感器的检测频率信号具有很好的放大效应,可以明显提高其检测灵敏度。     

石家庄西部太行山区景观格局时空变化

基于1987、2000年两期TM、ETM＋遥感数据资料,利用ArcGIS提取石家庄西部太行山区的景观类型转移矩阵,并结合重心模型,从时间和空间上对景观格局动态变化进行分析。结果表明：（1）1987～2000年,景观基质均为耕地,林地、草地、水域、建设用地和未利用土地作为斑块单元镶嵌其中;耕地、草地和水域的面积均为减少,其中草地面积减少量最大,主要向林地转化;林地、建设用地和未利用土地的面积均为增加,其中建设用地面积增加量最大,主要由未利用土地转化而来。（2）1987～2000年间,水域重心迁移距离最大;耕地、水域和建设用地的重心偏移方向为北偏西;林地和草地的重心偏移方向为北偏东;未利用土地的重心偏移方向为南偏西。     

SATB2 targeted by methylated miR‐34c‐5p suppresses proliferation and metastasis attenuating the epithelial‐mesenchymal transition in colorectal cancer

Objectives

SATB2 has been shown to be markedly reduced in colorectal cancer (CRC) tissues relative to paired normal controls; however, the mechanism behind remains not well understood. To investigate why SATB2 was down‐regulated in CRC, we attempted to analyse it from the angle of miRNA‐mRNA modulation.

Materials and methods

SATB2 expression was detected in CRC tissues using immunohistochemistry and verified using real‐time PCR on mRNA level, followed by analysis of clinicopathological significance of its expression. Metastatic variation of CRC cells was evaluated both in vivo and in vitro. To find out the potential miRNA that directly regulate the SATB2, luciferase reporter assay was performed following the bioinformatic prediction.

Results

SATB2 was confirmed to be closely linked with the metastasis and shorter overall survival of CRC in our own cases. Silencing of SATB2 was shown to be able to promote the metastatic ability of CRC cells in vivo, enhancing the epithelial‐mesenchymal transition (EMT). Mechanistically, miR‐34c‐5p was identified to be a novel miRNA that can directly modulate the SATB2. It turned out that the promoter of miR‐34c‐5p was methylated, which leads to the repression of miR‐34c‐5p in CRC. Treatment with 5‐Aza‐dC can reasonably and significantly restore the level of miR‐34c‐5p in CRC cells relative to control, thereby down‐regulating the SATB2.

Conclusions

Together, our study revealed that SATB2 targeted by methylated miR‐34c‐5p can suppress the metastasis, weakening the EMT in CRC.

     

基于多时相Landsat5/8影像的岷江汶川-都江堰段植被覆盖动态监测

植被覆盖度是衡量地表植被状况和指示生态环境变化的一个重要指标。基于像元二分模型,利用Landsat5/8遥感影像和DEM数据,对岷江汶川-都江堰段植被覆盖动态变化进行了监测,并结合高程、坡度和坡向数据,分析了汶川地震前后植被受损与恢复的空间动态格局变化。研究表明:植被覆盖总体良好,大部分区域的植被覆盖度均在中、高度以上,空间格局上呈现由汶川县东部、都江堰市西北部的龙门山区向两侧减少的总体趋势;地震造成植被受损面积达63808.7 hm~2,且集中分布于海拔567—4331 m、坡度26—51°的范围以及东坡、北坡、南坡和西坡;震后5a,植被恢复面积17786.47 hm~2,主要分布在海拔576—2180 m与3256—3793 m、坡度小于9°和26—51°以及东坡、东南坡和和南坡;高程和坡度对植被损毁与恢复的影响明显高于坡向。     

黄土高原丘陵区退耕还林地油松人工林碳储量及分配特征研究

以黄土高原丘陵区主要退耕还林树种油松为研究对象,对甘肃省庆阳市合水县采用样地调查与生物量实测方法,分析不同坡向（阳坡、阴坡）及退耕年限（退耕6年、9年和12年）油松人工林的乔木不同器官、灌草层、枯落物层和土壤层的碳含量,以及油松人工林乔木层、灌草层、枯落物层和土壤层碳储量及其分配特征,探讨甘肃黄土高原丘陵区生态林的固碳作用。结果表明:（1）油松不同器官碳含量为48.15%~53.90%,各器官碳含量大小为树干>叶>细枝>粗枝>根桩>粗根>树皮>大根>中根>小根>细根>球果;灌木层碳含量为茎>叶>根;草本层碳含量为地上部分>地下部分。（2）油松人工林的枯落物层碳含量为未分解层大于半分解层。（3）0~100 cm土壤层的碳含量随退耕年限增加而增大,随土壤深度的增加而下降;0~10 cm、10~20 cm土壤层不同坡向间碳含量差异显著。（4）阳坡和阴坡退耕6年、9年和12年油松林总碳储量分别为42.90、50.50、59.22 t·hm^-2和45.08、53.77、65.70 t·hm^-2。研究认为,黄土高原丘陵区阳坡和阴坡均适宜油松林发挥固碳效益,且阴坡要优于阳坡,是甘肃黄土高原丘陵区的理想树种。     

Spectrum structures and biological functions of 8-mers in the human genome

The spectra of k-mer frequencies can reveal the structures and evolution of genome sequences. We confirmed that the trimodal spectrum of 8-mers in human genome sequences is distinguished only by CG2, CG1 and CG0 8-mer sets, containing 2,1 or 0 CpG, respectively. This phenomenon is called independent selection law. The three types of CG 8-mers were considered as different functional elements. We conjectured that (1) nucleosome binding motifs are mainly characterized by CG1 8-mers and (2) the core structural units of CpG island sequences are predominantly characterized by CG2 8-mers. To validate our conjectures, nucleosome occupied sequences and CGI sequences were extracted, then the sequence parameters were constructed through the information of the three CG 8-mer sets respectively. ROC analysis showed that CG1 8-mers are more preference in nucleosome occupied segments (AUC > 0.7) and CG2 8-mers are more preference in CGI sequences (AUC > 0.99). This validates our conjecture in principle.     

The Acoustic Properties of Low Intensity Vocalizations Match Hearing Sensitivity in the Webbed-Toed Gecko,Gekko subpalmatus

The design of acoustic signals and hearing sensitivity in socially communicating species would normally be expected to closely match in order to minimize signal degradation and attenuation during signal propagation. Nevertheless, other factors such as sensory biases as well as morphological and physiological constraints may affect strict correspondence between signal features and hearing sensitivity. Thus study of the relationships between sender and receiver characteristics in species utilizing acoustic communication can provide information about how acoustic communication systems evolve. The genus Gekko includes species emitting high-amplitude vocalizations for long-range communication (loud callers) as well as species producing only low-amplitude vocalizations when in close contact with conspecifics (quiet callers) which have rarely been investigated. In order to investigate relationships between auditory physiology and the frequency characteristics of acoustic signals in a quiet caller, Gekko subpalmatus we measured the subjects’ vocal signal characteristics as well as auditory brainstem responses (ABRs) to assess auditory sensitivity. The results show that G. subpalmatus males emit low amplitude calls when encountering females, ranging in dominant frequency from 2.47 to 4.17 kHz with an average at 3.35 kHz. The auditory range with highest sensitivity closely matches the dominant frequency of the vocalizations. This correspondence is consistent with the notion that quiet and loud calling species are under similar selection pressures for matching auditory sensitivity with spectral characteristics of vocalizations.