RNA-guided CRISPR-Cas9 proteins have been widely used for genome editing, but their off-target activities limit broad application. The minimal Cas9 ortholog from Staphylococcus aureus (SaCas9) is commonly used for in vivo genome editing; however, no variant conferring high genome-wide specificity is available. Here, we report rationally engineered SaCas9 variants with highly specific genome-wide activity in human cells without compromising on-target efficiency. One engineered variant, referred to as SaCas9-HF, dramatically improved genome-wide targeting accuracy based on the genome-wide unbiased identification of double-stranded breaks enabled by sequencing (GUIDE-seq) method and targeted deep sequencing analyses. Among 15 tested human endogenous sites with the canonical NNGRRT protospacer adjacent motif (PAM), SaCas9-HF rendered no detectable off-target activities at 9 sites, minimal off-target activities at 6 sites, and comparable on-target efficiencies to those of wild-type SaCas9. Furthermore, among 4 known promiscuous targeting sites, SaCas9-HF profoundly reduced off-target activities compared with wild type. When delivered by an adeno-associated virus vector, SaCas9-HF also showed reduced off-target effects when targeting VEGFA in a human retinal pigmented epithelium cell line compared with wild type. Then, we further altered a previously described variant named KKH-SaCas9 that has a wider PAM recognition range. Similarly, the resulting KKH-HF remarkably reduced off-target activities and increased on- to off-target editing ratios. Our finding provides an alternative to wild-type SaCas9 for genome editing applications requiring exceptional genome-wide precision.

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With the research and the development of graphene-based materials, new sensors based on graphene compound materials are of great significance to scientific research and the consumer market. However, in the past ten years, due to the requirements of sensor accuracy, reliability, and durability, the development of new graphene sensors still faces many challenges in the future. Due to the special structure of graphene, the obtained characteristics can meet the requirements of high-performance sensors. Therefore, graphene materials have been applied in many innovative sensor materials in recent years. This paper introduces the important role and specific examples of sensors based on graphene and its base materials in biomedicine, photoelectrochemistry, flexible pressure, and other fields in recent years, and it puts forward the difficulties encountered in the application of graphene materials in sensors. Finally, the development direction of graphene sensors has been prospected. For the past two years of the COVID-19 epidemic, the detection of the virus sensor has been investigated. These new graphene sensors can complete signal detection based on accuracy and reliability, which provides a reference for researchers to select and manufacture sensor materials.

Ewing sarcomas are typified by EWSR1 fusion to ETS gene family members. Tumors with fusion partners other than ETS family members and atypical histologic features pose significant diagnostic challenges and controversies as to their classification. In this article, we report a tumor with EWSR1-NFATC2 fusion in the left femur of a 43-year-old man and with unusual morphologic features that resemble undifferentiated high-grade sarcoma. Analysis together with reported cases in the literature shows that tumors with EWSR1-NFATC2 exhibit distinctive clinicopathologic features, including predilection for young male adults, highly variable histology that varies from round cell tumors frequently associated with nuclear irregularity, short spindle cells with nuclear pleomorphism, to myoepithelial tumor-like with or without myxohyaline matrix. They show variable positivity to CD99, frequent expression of cytokeratins, and consistent high-level amplification of EWSR1-NFATC2 fusion gene with distinctive gene expression profile. These tumors thus deserve classification separate from Ewing sarcoma.

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As is well known, design is human oriented. Then, how can we realize human oriented when designing? The products are not made by way of designers' imagination; rather various design elements of the circumstance are integrated into the products so that the products are teemed with meanings. In modern society, the industrial products flood our life and various and mixed designs make us dazing. How to push modern design to move towards simplicity and reality is a question that deserves each designer's reflection. "Simplicity" refers to that a design must have explicit aim and solve the comparatively simplex problem; and the design language must be clear and express and the shape be unsophisticated. "Reality" refers to rational design concept and attitude and the maneuverability of the design.Keywords: Industrial design, Shape, Simplicity, Reality A traditional Chinese description of artworks is "being in possession of shape as well as bearing". This compliment means that the artwork is not only similar to the object described in shape, but also in bearing. In the shape design of modern industrial products, new shape images shall be pursued in accordance with the proper and intrinsic functions of the products. The symbolism connotation of the products shall not only reflect the intrinsic functions of the products, but also reflect the spiritual demands of the users on the products, which is the modernism connotation of "being in possession of shape as well as bearing" in industrial products.When conducting shape design for products, initially, the designers will usually feel shriveled. The reason for it is that the designers have not gained profound comprehension on this kind of products and do not have deep understanding and feeling towards the products; to put it in other words, the designers do not have profound understanding and master of the shape languages.The essential of shape is form. In our experience system, form that can be seen and touched is "reality form", while form that cannot be directly felt through vision and touch is "concept form".Sullivan mentioned many times his views on function and form. He said that "forms follow functions". This sentence must have great influence on Wright who had studied under the instruction of Sullivan for many years. Wright afterwards developed this sentence and put it into "functions are the same as forms", which shows his stand of functionalism. He believed that functions and forms could not be fully separated at all in designs. In the designs of Wright, a great many basic geometry graphics were adopted, such as squareness, rotundity and triangle, etc; in general design, gridding phalanx was adopted; together with various abstract detailed disposal, so a special decorating effect was reached. A strong characteristic of the design of Wright is that he endeavored to search harmonious elements between his designs and their surrounding natural environment, and tried to harmonize his designs with the nature and reach harmoniousness. His designs are organic compon...

Gene fusions are common drivers and therapeutic targets in cancers, but clinical-grade bioinformatics callers are lacking. Here we introduce a novel method SplitFusion, which is fast by leveraging BWA-MEM split alignments, can detect cryptic splice site fusions, and can infer frame-ness and exon-boundary alignments for functional prediction and minimizing false-positives. SplitFusion demonstrates superior sensitivity, specificity, accuracy and consumes minimal computing resources. In our study of 1,076 formalin-fixed paraffin-embedded lung cancer samples, SplitFusion detected not only common fusions (EML4 4.7%, ROS1 2.0% and RET 1.1%) with various partners, but also rare (KLC1-ALK, CD74-NRG1, and TPR-NTRK1) and novel (FGFR3-JAKMP1, CLIP2-BRAF, and ITPR2-ETV6) fusions. In 35 glioblastoma samples, SplitFusion-Target detected six (17%) EGFR vIII (exons 2-7 deletion) cases. Furthermore, we find that the EML4-ALK variant 3 is significantly associated with occurrence of multiple breakpoint-defined subclones, namely high intratumor heterogeneity. In conclusion, SplitFusion is well-suited for clinical use and for studying fusion-defined tumor heterogeneity.