新型冠状病毒纳米PCR快速检测方法的建立

新型冠状病毒(SARS-CoV-2)感染可导致致命性肺炎,且极具传染性。自2019年年末在我国出现以来,已在全球蔓延。为了建立一种灵敏、快速检测SARS-CoV-2的分子检测方法,本研究使用金纳米颗粒配合普通PCR检测方法,针对SARS-CoV-2核衣壳蛋白建立了SARS-CoV-2 NanoPCR新型分子检测方法。特异性结果显示,该方法对猪流行性腹泻病毒、牛冠状病毒、犬冠状病毒、水貂冠状病毒、猫传染性腹膜炎病毒均无交叉反应,表明该方法的特异性良好。敏感性结果显示,该方法的最低检测量为3.69×10⁴拷贝/μL,高于普通PCR最低检测量10倍,表明该方法的敏感性良好。使用建立的方法对3份临床样品进行检测,该方法与普通PCR方法结果一致,10倍稀释样品后,NanoPCR相比较普通PCR条带仍然具有较高辨识度。综上,本研究建立的灵敏、特异的NanoPCR方法可以对临床样品进行快速诊断和鉴定。     

毛竹实时荧光定量PCR内参基因的筛选及成花基因PheTFL1表达分析

通过qRT-PCR对毛竹相关成花基因PheTFL1的表达进行研究,为毛竹开花机理的研究提供理论依据.从毛竹UBC18、PP2A和EF1α等9个候选内参基因中筛选出在叶、幼嫩花序、花序轴、枝、竹青等11个组织器官中都稳定表达的PP2A用于毛竹PheTFL1基因qRT-PCR结果的校正.结果显示:PheTFL1基因在开花竹叶、枝和竹青中低丰度表达,与未开花竹差异不显著,但在花和花序轴中高丰度表达;在实生苗叶和根中高丰度表达,在实生苗茎中低丰度表达.PheTFL1基因在具有分生能力的幼嫩组织中高丰度表达,说明其不仅参与花发育的调控,还参与了分生组织生长的调控.     

Chemical synthesis and functional characterization of a new class of ceramide analogues as anti-cancer agents

Deregulation of ceramide metabolism is a hallmark of human cancer. Ceramide analogues thereby represent a new class of anti-cancer agents. We aimed at developing effective and low toxic ceramide analogues and synthesized a new class of ceramide analogues starting from l-threonine. Several analogues exhibit potent cytotoxicity against human cancer cells in vitro with IC₅₀ as low as 4.8?μM. These ceramide analogues decreased xIAP and Bcl-xL level and exhibited significant sensitization activity to overcome human cancer cell resistance to TRAIL, a cancer-selective agent that are being tested in human clinical trials. Furthermore, we determined that these ceramide analogues effectively suppress human cancer xenograft growth in vivo with no significant toxicity at the efficacious dose. Therefore, we have developed a simple and effective method to synthesize functional ceramide analogues using l-threonine as starting material and these analogues have the great potential to be further developed as anti-cancer agents in human cancer therapy.     

枯草芽孢杆菌中一种新型双向启动子的功能鉴定

本研究旨在通过转录组分析预测的方法,由地衣芽孢杆菌中筛选获得一种新型双向启动子,鉴定其启动强度。以已知强组成型启动子pShuttle-09为对照,检测其对克劳氏芽孢杆菌碱性蛋白酶基因的表达活性。成功构建了3种重组碱性蛋白酶表达载体及对应的工程菌株。在新型启动子pLA和其反向启动子pLB调控转录下,克劳氏芽孢杆菌碱性蛋白酶表达活性达到164 U/mL和111 U/mL。结果表明,pLA的启动强度明显高于pShuttle-09和pLB,pLA启动子与pLB启动子均可表达碱性蛋白酶。从而为枯草芽孢杆菌表达系统中异源基因的表达提供一个新的方向,也为原核生物中共同表达两种基因提供了新的思路。     

油葵苗期抗旱相关性状的QTL定位及候选基因筛选

干旱是限制向日葵生长发育的重要因素之一。为探究向日葵苗期抗旱性分子机制,该研究以向日葵K55与K58杂交构建的150个F₇重组自交系群体为材料,对其在正常浇水和干旱胁迫两种水分处理条件下的叶片相对电导率、叶绿素含量、叶面积、叶片相对含水量、根长进行表型测定,利用前期建立的SNP、SSR分子标记遗传连锁图谱,通过复合区间作图法对5个抗旱相关的性状进行QTL定位。结果表明：(1)共定位到向日葵QTL位点11个,其中正常浇水条件下5个,干旱胁迫条件下6个,表型贡献率为0.768%~7.547%,且5号连锁群上定位到的QTL位点最多(3个)。(2)QTL置信区间内共筛选到62个与干旱相关的候选基因,包括位于qLA 8 1上的rna23019、rna23004、rna22661、rna22193、rna23294、rna22783和位于qCC 13 1上的rna40140,这些基因可作为后续基因克隆及功能研究的重点候选基因。该研究结果为向日葵抗旱性研究及其遗传改良奠定了基础。     

Abacus Training Affects Math and Task Switching Abilities and Modulates Their Relationships in Chinese Children

Our previous work demonstrated that abacus-based mental calculation (AMC), a traditional Chinese calculation method, could help children improve their math abilities (e.g. basic arithmetical ability) and executive function (e.g. working memory). This study further examined the effects of long-term AMC training on math ability in visual-spatial domain and the task switching component of executive function. More importantly, this study investigated whether AMC training modulated the relationship between math abilities and task switching. The participants were seventy 7-year-old children who were randomly assigned into AMC and control groups at primary school entry. Children in AMC group received 2-hour AMC training every week since primary school entry. On the contrary, children in the control group had never received any AMC training. Math and task switching abilities were measured one year and three years respectively after AMC training began. The results showed that AMC children performed better than their peers on math abilities in arithmetical and visual-spatial domains. In addition, AMC group responded faster than control group in the switching task, while no group difference was found in switch cost. Most interestingly, group difference was present in the relationships between math abilities and switch cost. These results implied the effect of AMC training on math abilities as well as its relationship with executive function.     

Lentivirus-mediated overexpression of TGF-β inducible early gene 1 inhibits SW1990 pancreatic cancer cell growth

TIEG1 (TGF-β inducible early gene 1) plays a significant role in regulating cell proliferation and apoptosis in various cell types. Previous studies have shown a close relationship between the expression level of TIEG1 and various cancers, including breast, prostate, colorectal and pancreatic cancer. In this study, we up-regulated the gene expression of TIEG1 in SW1990 pancreatic cancer cell line by a lentivirus transfection system and investigated its potential as a therapeutic target for pancreatic cancer. The results showed that lentivirus-mediated overexpression of TIEG1 gene inhibited human pancreatic cancer SW1990 cell proliferation and caused the cell cycle arrest at the G1-phase in vitro. SW1990 cells transduced with lenti-TIEG1 showed significant inhibition of colony formation and cancer cell growth in 3-D culture model. Moreover, overexpression of TIEG1 gene significantly slowed the growth of SW1990 xenografts in nude mice. Taken together, these data provided evidence that overexpression of TIEG1 gene by a lentivirus transfection system led to suppressed human pancreatic cancer cell growth and might therefore be a feasible approach in the clinical management of pancreatic cancer.     

A porcine circovirus type 2 (PCV2) mutant with 234 amino acids in capsid protein showed more virulence in vivo, compared with classical PCV2a/b strain

Background

Porcine circovirus type 2 (PCV2) is considered to be the primary causative agent of postweaning multisystemic wasting syndrome (PMWS), which has become a serious economic problem for the swine industry worldwide. The major genotypes, PCV2a and PCV2b, are highly prevalent in the pig population and are present worldwide. However, another newly emerging PCV2b genotype mutant, which has a mutation in its ORF2-encoded capsid protein, has been sporadically present in China, as well as in other countries. It is therefore important to determine the relative virulence of the newly emerging PCV2b genotype mutant, compared with the existing PCV2a and PCV2b genotypes, and to investigate whether the newly emerging mutant virus induces more severe illness.

Methodology/Principal Findings

Twenty healthy, 30-day-old, commercial piglets served as controls or were challenged with PCV2a, PCV2b and the newly emerging mutant virus. A series of indexes representing different parameters were adopted to evaluate virulence, including clinical signs, serological detection, viral load and distribution, changes in immune cell subsets in the peripheral blood, and evaluation of pathological lesions. The newly emerging PCV2 mutant demonstrated more severe signs compatible with PMWS, characterized by wasting, coughing, dyspnea, diarrhea, rough hair-coat and depression. Moreover, the pathological lesions and viremia, as well as the viral loads in lymph nodes, tonsils and spleen, were significantly more severe (P<0.05) for piglets challenged with the newly emerging mutant compared with those in the groups challenged with PCV2a and PCV2b. In addition, a significantly lower average daily weight gain (P<0.05) was recorded in the group challenged with the newly emerging PCV2 mutant than in the groups challenged with the prevailing PCV2a and PCV2b.

Conclusions

This is believed to be the first report to confirm the enhanced virulence of the newly emerging PCV2 mutant in vivo.