Increasing yield potential through manipulating of an ARE1 ortholog related to nitrogen use efficiency in wheat by CRISPR/Cas9.

peer reviewedWheat (Triticum aestivum L.) is a staple food crop consumed by more than 30% of world population. Nitrogen (N) fertilizer has been applied broadly in agriculture practice to improve wheat yield to meet the growing demands for food production. However, undue N fertilizer application and the low N use efficiency (NUE) of modern wheat varieties are aggravating environmental pollution and ecological deterioration. Under nitrogen-limiting conditions, the rice (Oryza sativa) abnormal cytokinin response1 repressor1 (are1) mutant exhibits increased NUE, delayed senescence and consequently, increased grain yield. However, the function of ARE1 ortholog in wheat remains unknown. Here, we isolated and characterized three TaARE1 homoeologs from the elite Chinese winter wheat cultivar ZhengMai 7698. We then used CRISPR/Cas9-mediated targeted mutagenesis to generate a series of transgene-free mutant lines either with partial or triple-null taare1 alleles. All transgene-free mutant lines showed enhanced tolerance to N starvation, and showed delayed senescence and increased grain yield in field conditions. In particular, the AABBdd and aabbDD mutant lines exhibited delayed senescence and significantly increased grain yield without growth defects compared to the wild-type control. Together, our results underscore the potential to manipulate ARE1 orthologs through gene editing for breeding of high-yield wheat as well as other cereal crops with improved NUE

Study on the accessibility and affordability of 50 drugs in Wuhan based on the WHO/HAI standardization method

ObjectiveTo understand the availability and affordability of essential drugs in Wuhan since the implementation of the national essential medicine system, and to provide a basis for the subsequent formulation and improvement of related policies.MethodsUsing the standard survey method jointly developed by the WHO and Health Action International (HAI), a sample of 50 essential drugs was selected to investigate and evaluate their availability and affordability in public medical and health institutions and social retail pharmacies in Wuhan, using six diseases with high clinical morbidity as the targets.ResultsThe availability of the original drug and the lowest-priced generic drug in public hospitals is 26.4 and 42.47% respectively, and that in retail pharmacies is 26.8 and 54.4% respectively. The median price ratio of the original drug and the lowest-priced generic drug is 28.71 and 2.23 respectively in public hospitals, and 29.24 and 3.59 respectively in retail pharmacies; In addition to individual drugs, such as omeprazole, others are affordable. The availability of essential drugs in public hospitals in Wuhan is lower than that in social retail pharmacies, and the availability of the lowest-priced generic drugs is much higher than that of original drugs.ConclusionThe availability of essential drugs in public hospitals in Wuhan is lower than that in social retail pharmacies, and the availability of the lowest-priced generic drugs is much higher than that of original drugs. The price of the original drug is much higher than the international reference price; The price of medicines in public hospitals is lower than that in retail pharmacies;the overall condition of affordability is good, but there is a big gap between the affordability levels of original drugs and generic drugs, and the affordability of original drugs is relatively poor. It is recommended to adjust the relevant policies according to the actual situation of Wuhan city itself, moderately ensure the supply of original drugs, improve the price transparency of retail pharmacies, and ensure that the basic drug needs of the public are met

Role of Socially Assistive Robots in Reducing Anxiety and Preserving Autonomy in Children

Anxiety in children is gradually becoming a problem that needs to be addressed urgently. Socially Assistive Robots (SARs) have shown great potential in anxiety treatment among adults and elders. However, the application on childhood anxiety has scarcely been tested. Autonomy is also an influential factor of psychological therapy sessions in children. This study, using state anxiety as a kick-off point, aims to investigate the effectiveness of SARs and the role of autonomy in reducing children's anxiety by using Progressive Muscle Relaxation (PMR). Participants were 69 Chinese children aged from 10 to 12. We found that SARs significantly reduced state anxiety levels in all conditions. But no difference between each level of intervention and autonomy was found. Further research on various perspectives is suggested

The influencing factors of biomedical R&D cooperation in three major urban agglomerations of China based on cooperative patents.

Due to the particularity of biomedical industry, it has become necessary for biomedical enterprises to seek innovative research and development (R&D) cooperation to maintain advanced technologies and products in multiple fields. Under such circumstance, the biomedical industry has gradually formed a certain cluster to promote the development of the industry. So far, the biomedical industry cluster has formed in China, mainly within the Yangtze River Delta, Pearl River Delta, and Beijing-Tianjin-Hebei three urban agglomerations. Within the industrial clusters, the frequency of innovation cooperation among enterprises, universities, research institutions, and other relevant organizations in the biomedical area is high, and the capacity for innovation cooperation is strong as well. This paper used the representative cross-section data of cooperative patents from the medical science and technology patent database of China National Knowledge Infrastructure (CNKI), researching the R&D cooperation within the three major urban agglomerations in China from 2008 to 2016 (Yangtze River Delta Urban Agglomeration, Pearl River Delta Urban Agglomeration, Beijing-Tianjin-Hebei Urban Agglomeration) on total 36 cities' spatial pattern characteristics of biomedical cooperation and the influencing factors. The spatial interaction model was used to study the spatial, economic, political, and R&D influencing factors of cross-city cooperation. The degree of aggregation showed that cross-city R&D cooperation mainly occurred in well-developed and central cities of urban agglomerations. Econometric results revealed that spatial, economic, political, and R&D bias factors did have a significant impact on the frequency of biomedical R&D cooperation across cities

Synthesis-dependent repair of Cpf1-induced double strand DNA breaks enables targeted gene replacement in rice.

The recently developed CRISPR (clustered regularly interspaced short palindromic repeats)/Cpf1 system expands the range of genome editing and is emerging as an alternative powerful tool for both plant functional genomics and crop improvement. Cpf1-CRISPR RNA (crRNA) produces double strand DNA breaks (DSBs) with long 5'-protruding ends, which may facilitate the pairing and insertion of repair templates through homology-directed repair (HDR) for targeted gene replacement and introduction of the desired DNA elements at specific gene loci for crop improvement. However, the potential mechanism underlying HDR of DSBs generated by Cpf1-crRNA remains to be investigated, and the inherent low efficiency of HDR and poor availability of exogenous donor DNA as repair templates strongly impede the use of HDR for precise genome editing in crop plants. Here, we provide evidence of synthesis-dependent repair of Cpf1-induced DSBs, which enables us precisely to replace the wild-type ALS gene with the intended mutant version that carries two discrete point mutations conferring herbicide resistance to rice plants. Our observation that the donor repair template (DRT) with only the left homologous arm is sufficient for precise targeted allele replacement offers a better understanding of the mechanism underlying HDR in plants, and greatly simplifies the design of DRTs for precision genome editing in crop improvement

Heterologous expression and antitumor activity analysis of syringolin from Pseudomonas syringae pv. syringae B728a

Abstract Background Syringolin, synthesized by a mixed non-ribosomal peptide synthetase/polyketide synthetase in Pseudomonas syringae pv. syringae (Pss) B728a, is a novel eukaryotic proteasome inhibitor. Meanwhile, directly modifying large fragments in the PKS/NRPS gene cluster through traditional DNA engineering techniques is very difficult. In this study, we directly cloned the syl gene cluster from Pss B301D-R via Red/ET recombineering to effectively express syringolin in heterologous hosts. Results A 22 kb genomic fragment containing the sylA–sylE gene cluster was cloned into the pASK vector, and the obtained recombinant plasmid was transferred into Streptomyces coelicolor and Streptomyces lividans for the heterologous expression of syringolin. Transcriptional levels of recombinant syl gene in S. coelicolor M145 and S. lividans TK24 were evaluated via RT-PCR and the production of syringolin compounds was detected via LC–MS analysis. The extracts of the engineered bacteria showed cytotoxic activity to B16, 4T1, Meth-A, and HeLa tumor cells. It is noteworthy that the syringolin displayed anticancer activity against C57BL/6 mice with B16 murine melanoma tumor cells. Together, our results herein demonstrate the potential of syrinolin as effective antitumor agent that can treat various cancers without apparent adverse effects. Conclusions This present study is the first to report the heterologous expression of the entire syl gene cluster in Streptomyces strains and the successful expression of syringolin in both S. coelicolor M145 and S. lividans TK24. Syringolin derivatives demonstrated high cytotoxicity in vitro and in vivo. Hence, this paper provided an important foundation for the discovery and production of new antitumor compounds

Estimation results of the negative binomial spatial interaction model, based on common patents among cities in Beijing-Tianjin-Hebei and Pearl River Delta urban agglomerations (2018–2014).

Estimation results of the negative binomial spatial interaction model, based on common patents among cities in Beijing-Tianjin-Hebei and Pearl River Delta urban agglomerations (2018–2014).</p