Expression,purification, and breast cancer cell inhibiting effect of recombinant human lactoferrin C-lobe

Lactoferrin (LTF), a multifunctional glycoprotein of the transferrin family mainly found in exotic secretions in mammals, is an important defense molecule against not only microbial invasion but also tumors. It folds into two globular domains (N- and C-lobes) each containing an iron-binding site. The cationic antimicrobial peptide in N-lobe is known to exert anti-tumor effect via a non-receptor-mediated pathway. However, whether LTF C-lobe also contributes to its anti-tumor activity remains to be investigated. In this study, a human LTF fragment (amino acid residues 343–682) covering the C-lobe was expressed with a histidine tag in E. coli and the purified polypeptide refolded through a series of buffer changing procedure. The resultant recombinant protein caused significant growth arrest of breast carcinoma cells MDA-MB-231 in a dose- and time-dependent manner, evidently via induction of apoptosis of the cell. Our data suggest a positive role for the C-lobe of human LTF in controlling tumors in vitro.     

Comparison of droplet digital PCR to real-time PCR for quantification of hepatitis B virus DNA

Quantitative real-time PCR (qPCR) has been widely implemented for clinical hepatitis B viral load testing, but a lack of standardization and relatively poor precision hinder its usefulness. Droplet digital PCR (ddPCR) is a promising tool that offers high precision and direct quantification. In this study, we compared the ddPCR QX100 platform by Bio-Rad with the CFX384 Touch Real-Time PCR Detection System (Bio-Rad, USA) to detect serial plasmid DNA dilutions of known concentrations as well as HBV DNA extracted from patient serum samples. Both methods showed a high degree of linearity and quantitative correlation. However, ddPCR assays generated more reproducible results and detected lower copy numbers than qPCR assays. Patient sample quantifications by ddPCR and qPCR were highly agreeable based on the Bland–Altman analysis. Collectively, our findings demonstrate that ddPCR offers improved analytical sensitivity and specificity for HBV measurements and is suitable for clinical HBV detection.     

Molecular Identification and Diversity of ‘Candidatus Phytoplasma solani’ Associated with Red‐leaf Disease of Salvia miltiorrhiza in China

Reddening disease has recently been threatening Salvia miltiorrhiza in China, ranging from 30 to 50%. The main symptoms observed, such as plant stunting, inflorescence malformation, leaf reddening, fibrous roots browning, skin blackening and eventually root rot, are typically associated with phytoplasma infection. The presence of phytoplasmas was demonstrated through phytoplasma‐specific PCR, with the expected amplification (1.8 kb) from symptomatic S. miltiorrhiza plants from Shangluo, Shangzhou and Luonan fields in Shaanxi Province of China. The sequences of 16S rRNA, tuf, secY and vmp1 genes amplified from LN‐1 phytoplasma shared the closest homologies of 99%, 100%, 99% and 98% with those of the reference strain Candidatus Phytoplasma solani (subgroup 16SrXII‐A), respectively. The phylogenetic trees showed that LN‐1 phytoplasma clustered with the members of 16SrXII‐A group, including Ca. P. solani. Computer‐simulated restriction fragment length polymorphism analysis further supported this classification. Diversity analysis showed that all ‘Ca. P. solani’ strains identified from the three different regions examined shared 100% identical 16S rRNA, tuf, secY and vmp1 nucleotide sequences. To the best of our knowledge, this is the first report of phytoplasma infecting the medicinal plant of S. miltiorrhiza. The results demonstrate that ‘Ca. P. solani’ is the presumptive aetiological agent of S. miltiorrhiza reddening disease in China.     

Microalgal industry in China: challenges and prospects

Over the past 15 years, China has become the major producer of microalgal biomass in the world. Spirulina (Arthrospira) is the largest microalgal product by tonnage and value, followed by Chlorella, Dunaliella, and Haematococcus, the four main microalgae grown commercially. China’s production is estimated at about two-thirds of global microalgae biomass of which roughly 90 % is sold for human consumption as human nutritional products (‘nutraceuticals’), with smaller markets in animal feeds mainly for marine aquaculture. Research is also ongoing in China, as in the rest of the world, for other high-value as well as commodity microalgal products, from pharmaceuticals to biofuels and CO₂ capture and utilization. This paper briefly reviews the main challenges and potential solutions for expanding commercial microalgae production in China and the markets for microalgae products. The Chinese Microalgae Industry Alliance (CMIA), a network founded by Chinese microalgae researchers and commercial enterprises, supports this industry by promoting improved safety and quality standards, and advancement of technologies that can innovate and increase the markets for microalgal products. Microalgae are a growing source of human nutritional products and could become a future source of sustainable commodities, from foods and feeds, to, possibly, fuels and fertilizers.     

Spectral feature differences between shrub and grass communities and shrub coverage retrieval in shrub-encroached grassland in Xianghuang Banner,Nei Mongol,China北大核心CSCD

Aims: Shrub-encroached grassland has become an important vegetation type in China's arid and semi-arid region. Our study objective is to explore the spectral features of shrub and grass communities, as well as their empirical relationships with shrub coverage. The quantitative estimation of shrub cover based on medium-resolution Landsat satellite imagery provides the practical basis for long term retrieval of large areas of shrub expansion in the grassland region. Methods: Linear models and Multiple Endmember Spectral Analysis Model (MESMA) based on medium resolution Landsat satellite imagery were developed to quantify the shrub coverage in a shrub-encroached grassland region in Xianghuang Banner, Nei Mongol using the spectral features and their seasonal differences between the shrub and grass communities. Important findings: Compared to Leymus chinensis and Stipa krylovii dominated grass communities, Caragana microphylla community had a higher normalized difference vegetation index (NDVI), modified red edge normalized difference vegetation index (mNDVI705), and red edge slope. The red edge position of C. microphylla community shifted to longer wavelengths. The average and the maximum shrub coverage was 13% and 25%, respectively, in the shrub-encroached grassland based on both models. The correlation coefficient of determination (R2) and root mean square error (RMSE) of the linear model was 0.31 and 0.05, respectively. We found that the linear model based on seasonal differences of shrub and grass community was more suitable for retrieving shrub coverage in the study area from medium resolution imagery than the MESMA model that is based on mid-summer images.     

Reconstitution of CO2 Regulation of SLAC1 Anion Channel and Function of CO2-Permeable PIP2;1 Aquaporin as CARBONIC ANHYDRASE4 Interactor

Dark respiration causes an increase in leaf CO₂ concentration (Ci), and the continuing increases in atmospheric [CO₂] further increases Ci. Elevated leaf CO₂ concentration causes stomatal pores to close. Here, we demonstrate that high intracellular CO₂/HCO₃⁻ enhances currents mediated by the Arabidopsis thaliana guard cell S-type anion channel SLAC1 upon coexpression of any one of the Arabidopsis protein kinases OST1, CPK6, or CPK23 in Xenopus laevis oocytes. Split-ubiquitin screening identified the PIP2;1 aquaporin as an interactor of the βCA4 carbonic anhydrase, which was confirmed in split luciferase, bimolecular fluorescence complementation, and coimmunoprecipitation experiments. PIP2;1 exhibited CO₂ permeability. Mutation of PIP2;1 in planta alone was insufficient to impair CO₂- and abscisic acid-induced stomatal closing, likely due to redundancy. Interestingly, coexpression of βCA4 and PIP2;1 with OST1-SLAC1 or CPK6/23-SLAC1 in oocytes enabled extracellular CO₂ enhancement of SLAC1 anion channel activity. An inactive PIP2;1 point mutation was identified that abrogated water and CO₂ permeability and extracellular CO₂ regulation of SLAC1 activity. These findings identify the CO₂-permeable PIP2;1 as key interactor of βCA4 and demonstrate functional reconstitution of extracellular CO₂ signaling to ion channel regulation upon coexpression of PIP2;1, βCA4, SLAC1, and protein kinases. These data further implicate SLAC1 as a bicarbonate-responsive protein contributing to CO₂ regulation of S-type anion channels.     

Genome‐wide association study of callus induction variation to explore the callus formation mechanism of rice

Rice(Oryza sativa) is one of the most widely cultivated food crops, worldwide. Tissue culture is extensively used in rice breeding and functional genome research. The ability to induce callus determines whether a particular rice variety can be subjected to tissue culture and Agrobacterium-mediated transformation. Over the past two decades, many quantitative trait loci(QTLs)related to callus induction traits have been identified;however, individual genes associated with rice callus induction have not been reported. In this study, we characterized three callus-induction traits in a global collection of 510 rice accessions. A genome-wide association study of the rice population in its entirety as well as subpopulations revealed 21 significant loci located in rice callus induction QTLs. We identified three candidate callus induction genes, namely CRL1, Os BMM1, and Os SET1, which Rese are orthologs of Arabidopsis LBD17/LBD29, BBM, and SWN,respectively, which are known to affect callus formation.Furthermore, we predicted that 14 candidate genes might be involved in rice callus induction and showed that RNA interference(RNAi)-mediated disruption of Os IAA10 inhibited callus formation on tissue culture medium.Embryo growth in the Os IAA10 RNAi line was not inhibited by synthetic auxin(2,4-D) treatment, suggesting that Os IAA10 may perceive auxin and activate the expression of downstream genes, such as CRL1, to induce callus formation. The significant loci and candidate genes identified here may provide insight into the mechanism underlying callus formation in rice.     

Plant peroxisomes:Small organelles with versatility and complexity

<正>Peroxisomes are small, highly dynamic, and multifunctional organelles in eukaryotes. Essential to plant survival, peroxisomes house various crucial metabolic activities, such as degradation of hydrogen peroxide(H_2O_2), lipid metabolism, photorespiration, and hormone biosynthesis and catabolism, and remodel their proteome in response to developmental and environmental changes(Hu et al. 2012; Pan and Hu 2018).     

小鼠胸主动脉血管管壁结构和平滑肌细胞表型的年龄变化

目的:通过研究小鼠胸主动脉血管直径和管壁厚度以及平滑肌细胞表型标志物波形蛋白(Vimentin)和平滑肌细胞肌动蛋白(α-SM-actin)的表达,探讨胸主动脉在发育、成熟和老化过程中血管管壁结构和平滑肌细胞表型的变化。方法:将24只昆明小鼠按年龄分成4组:3天组、3个月组、6个月组和16个月组。采用HE染色和免疫荧光术分别观察胸主动脉血管管壁结构以及Vimentin和α-SM-actin的表达变化。结果:随着年龄的增长,胸主动脉血管直径和管壁厚度增加,以16个月小鼠胸主动脉的直径最大和管壁最厚,但细胞密度减少。3天小鼠胸主动脉的Vimentin表达较高,6个月表达下降,16个月重新上调;而α-SM-actin在3天小鼠的胸主动脉表达较低,6个月表达增加,16个月表达出现下调,其差异均有统计学意义(P0.05)。结论:小鼠胸主动脉的直径和管壁的厚度随年龄的增长而增加,而细胞的密度则随着年龄增加而减少;平滑肌细胞的表型从幼龄时的合成表型转变为收缩表型,老龄时又转变为合成表型。