昆虫细胞（Sf21）悬浮培养过程中生长限制性基质间歇补加技术的 …

基于ｒ２１昆虫细胞在浮过程中所表现出的生长代谢特征,提出以培养液中残糖浓度作为控制参数,并利用限制性基质（葡萄糖和蛋白水解物）的间歇补加技术调控细胞生长的方案。实际控制表明：与批培养相比,Ｓ１ｆ２１细胞在两种具代表性的昆虫水解物）的间歇补加调控细胞生长的方案。实际控制表明：与批培养相比,Ｓｆ２１细胞在两种具代表性的昆虫细胞培养基（ＩＰＬ－４１和ＴＣ－１００）中的生长期和稳定期部都到有效的延长。ＴＣ     

应用引物延伸预扩增提高微量DNA拷贝数

采用引物延伸预扩增方法 ,可普遍提高微量模板DNA的拷贝数 ,便于进行基因分析时克服标本量少、来源困难的制约。采用常规扩增、检测 2 4 8bp的DYZ1片段体系为观察对象 ,其最小模板量需 1.5ng/2 0 μl体系。以 15个碱基随机寡核苷酸为引物 ,对最小模板量进行预扩增 ,再以其产物 1/10为模板 ,特异扩增DYZ1片段。进行相对定量分析 ,判断原模板DNA拷贝数增加的程度。结果 1.5ng男性DNA经随机扩增后 ,此DYZ1片段拷贝数增加了 10倍以上 ,大大地提高了特异DNA片段扩增的模板量。表明经随机引物延伸预扩增后 ,微量标本DNA片段拷贝数获得普遍提高 ,增加了微量DNA扩增的敏感度     

粘虫成虫在气流场中飞行行为的观察研究

通过改进悬吊测飞技术、室内风洞和野外雷达相结合的观测方法,研究了粘虫在气流场中的飞行行为特征及其与气流的关系。直筒风洞自由飞行观测的结果表明,粘虫蛾对气流有明显行为反应,表现为头部迎风起飞和迎风飞行的特性;在3.0～5.5 m/s风速下,有92%～94% 的个体可一次逆风飞行通过2 m长的风洞;当风速≥6.0 m/s时,有71.9%的蛾子沿螺旋状的飞行轨迹逆风通过风洞。环形风洞悬吊飞行测试的结果表明,粘虫可逆风飞行的最大风速为7.2 m/s;在风速≤4 m/s条件下,90%以上个体头部迎风飞行或头部朝向与风向成一定的夹角,侧逆风飞行。 雷达观测发现粘虫在空中迁飞过程中具有成层现象,并有较强的秋季回迁定向行为,其头部总是朝向西南;迁飞的最终位移与风向及风速大小有关,迁飞位移速度是飞行速度与风速的矢量和。     

Gene Presence/Absence Variation analysis of coronavirus family displays its pan-genomic diversity

SARS-CoV-2 belongs to the coronavirus family. Comparing genomic features of viral genomes of coronavirus family can improve our understanding about SARS-CoV-2. Here we present the first pan-genome analysis of 3,932 whole genomes of 101 species out of 4 genera from the coronavirus family. We found that a total of 181 genes in the pan-genome of coronavirus family, among which only 3 genes, the S gene, M gene and N gene, are highly conserved. We also constructed a pan-genome from 23,539 whole genomes of SARS-CoV-2. There are 13 genes in total in the SARS-CoV-2 pan-genome. All of the 13 genes are core genes for SARS-CoV-2. The pan-genome of coronaviruses shows a lower level of diversity than the pan-genomes of other RNA viruses, which contain no core gene. The three highly conserved genes in coronavirus family, which are also core genes in SARS-CoV-2 pan-genome, could be potential targets in developing nucleic acid diagnostic reagents with a decreased possibility of cross-reaction with other coronavirus species.     

Low-density lipoprotein receptor-related protein 1 is an essential receptor for trichosanthin in 2 choriocarcinoma cell lines

Type-I ribosome-inactivating protein-trichosanthin (TCS) exhibits selective cytotoxicity toward different types of cells. It is believed that the cytotoxicity results from the inhibition of ribosomes to decrease protein synthesis, thereby indicating that there are specific mechanisms for TCS entry into target cells to reach the ribosomes. Low-density lipoprotein (LDL) receptor-related protein 1 (LRP1) is a large scavenger receptor that is responsible for the binding and endocytosis of diverse biological ligands on the cell surface. In this study, we demonstrated that 2 choriocarcinoma cell lines can significantly bind and internalize TCS. In contrast, Hela cell line displayed no obvious TCS binding and endocytosis. Furthermore LRP1 gene silencing in JAR and BeWo cell lines blocked TCS binding; TCS could also interact with LRP1.The results of our study established that LRP1 was a major receptor for phagocytosis of TCS in JAR and BeWo cell lines and might be the molecular basis of TCS abortificient and anti-choriocarcinoma activity.     

Ultrasound-Assisted Extraction of Water-Soluble Polysaccharides from the Fruit of Acanthopanaxbrachypus: Physicochemical,Structural and Functional Properties

The ultrasound-assisted extraction (UAE) parameters of total water-soluble polysaccharides (TABPs) from Acanthopanaxbrachypus fruit were optimized by Box-Behnken design (BBD) and response surface methodology (RSM). Physicochemical, structural, and functional properties of TABPs were investigated by chemical analysis, inductively coupled plasma optical emission spectrometry (ICP-OES), high performance liquid chromatography (HPLC), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA), water-holding capacity (WHC), oil-holding capacity (OHC), emulsion capacity (EC), emulsion stability (ES), as well as DPPH^. and ABTS^.+scavenging assays. The results showed that the maximal UAE-yield of TABPs was 3.81±0.18 % under the optimal conditions (ultrasonic power 325 W, extraction temperature 47 °C, extraction time 22 min). TABPs was rich in some beneficial element (Mg, K, Fe, Zn and Na) but little in harmful elements (Hg, Cd, As and Pb), and displayed rough surface with flake-like features and large dents, contained 93.89±0.08 % of total carbohydrate with more different monosaccharides including glucose, galactose, rhamnose, arabinose, mannose, xylose, and uronic acid in a molar ratio of 8.83 : 7.90 : 4.74 : 4.55 : 2.80 : 2.39 : 1.00, respectively. TABPs exhibited broad weight distribution (11.2–133.5 kDa), excellent thermal stability (>280 °C), WHC (0.61±0.08 g water/g sample) and OHC (4.53±0.12 g oil/g sample), as well as higher EC (43.75±1.23 %) and ES (38.32±1.50 %). Furthermore, TABPs also displayed remarkable scavenging activities on DPPH^. and ABTS^.+ in vitro. These findings provide a scientific basis for the applications of TABPs in functional additives for food, medicine, and cosmetics.     

The Evolutionary Dynamics of Genetic Incompatibilities Introduced by Duplicated Genes in Arabidopsis thaliana

Although gene duplications provide genetic backup and allow genomic changes under relaxed selection, they may potentially limit gene flow. When different copies of a duplicated gene are pseudofunctionalized in different genotypes, genetic incompatibilities can arise in their hybrid offspring. Although such cases have been reported after manual crosses, it remains unclear whether they occur in nature and how they affect natural populations. Here, we identified four duplicated-gene based incompatibilities including one previously not reported within an artificial Arabidopsis intercross population. Unexpectedly, however, for each of the genetic incompatibilities we also identified the incompatible alleles in natural populations based on the genomes of 1,135 Arabidopsis accessions published by the 1001 Genomes Project. Using the presence of incompatible allele combinations as phenotypes for GWAS, we mapped genomic regions that included additional gene copies which likely rescue the genetic incompatibility. Reconstructing the geographic origins and evolutionary trajectories of the individual alleles suggested that incompatible alleles frequently coexist, even in geographically closed regions, and that their effects can be overcome by additional gene copies collectively shaping the evolutionary dynamics of duplicated genes during population history.     

Long-term clinical results of autologous infusion of mobilized adult bone marrow derived CD34+ cells in patients with chronic liver disease

Abstract.  Evidence is growing in support of the role of stem cells as an attractive alternative in treatment of liver diseases. Recently, we have demonstrated the feasibility and safety of infusing CD34⁺ adult stem cells; this was performed on five patients with chronic liver disease. Here, we present the results of long-term follow-up of these patients. Between 1 × 10⁶ and 2 × 10⁸ CD34⁺ cells were isolated and injected into the portal vein or hepatic artery. The patients were monitored for side effects, toxicity and changes in clinical, haematological and biochemical parameters; they were followed up for 12–18 months. All patients tolerated the treatment protocol well without any complications or side effects related to the procedure, also there were no side effects noted on long-term follow-up. Four patients showed an initial improvement in serum bilirubin level, which was maintained for up to 6 months. There was marginal increase in serum bilirubin in three of the patients at 12 months, while the fourth patient's serum bilirubin increased only at 18 months post-infusion. Computed tomography scan and serum α-foetoprotein monitoring showed absence of focal lesions. The study indicated that the stem cell product used was safe in the short and over long term, by absence of tumour formation. The investigation also illustrated that the beneficial effect seemed to last for around 12 months. This trial shows that stem cell therapy may have potential as a possible future therapeutic protocol in liver regeneration.