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Improving Plant Genome Editing with High-Fidelity xCas9 and Non-canonical PAM-Targeting Cas9-ng

Molecular Plant 2019年第7期12卷 1027-1036页

作者： Zhaohui Zhong simon Sretenovic Qiurong Ren Lijia Yang Yu Bao Caiyan Qi Mingzhu Yuan Yao He Shishi Liu Xiaopei Liu Jiaheng Wang Lan Huang Yan Wang Dibin Baby David Wang Tao Zhang Yiping Qi Yong Zhang Department of Biotechnology School of Life Sciences and Technology Center for Informational Biology University of 曰ectronic Science and Technology of China Room 216 Main Building No. 4 Section 2 North Jianshe Road Chengdu 610054 China Department of Plant Science and Landscape Architecture University of Maryland College Park MD 20742 USA Jiangsu Key Laboratory of Crop Genetics and Physiology Co-Innovation Center for Modern Production Technology of Grain Crops Key Laboratory of Plant Functional Genomics of the Ministry of Education Yangzhou University Yangzhou 225009 China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China Yangzhou University Yangzhou 225009 China Indian Institute of Science Education and Research Timpati Andhra Pradesh 517507 India Seven Lakes High School Katy TX 77494 USA Institute for Bioscience and Biotechnology Research University of Maryland Rockville MD 20850 USA

Two recently engineered SpCas9 variants, namely xCas9 and Cas9-ng, show promising potential in improving targeting specificity and broadening the targeting range. In this study, we evaluated these Cas9 variants in the model and crop plant, rice. We first tested xCas9-3.7, the most effective xCas9 variant in mammalian cells, for targeted mutagenesis at 16 possible ngN PAM (protospacer adjacent motif) combinations in duplicates. xCas9 exhibited nearly equivalent editing efficiency to wild-type Cas9 (Cas9-WT) at most canonical ngG PAM sites tested, whereas it showed limited activity at non-canonical ngH (H = A, C, T) PAM sites. High editing efficiency of xCas9 at ngG PAMs was further demonstrated with C to T base editing by both rAPOBECI and PmCDAI cytidine deaminases. With mismatched sgRNAs, we found that xCas9 had improved targeting specificity over the Cas9-WT. Furthermore, we tested two Cas9-ng variants, Cas9-ngv1 and Cas9-ng, for targeting ngN PAMs. Both Cas9-ng variants showed higher editing efficiency at most non-canonical ng PAM sites tested, and enabled much more efficient editing than xCas9 at AT-rich PAM sites such as GAT, GAA, and CAA. Nevertheless, we found that Cas9-ng variants showed significant reduced activity at the canonical ngG PAM sites. In stable transgenic rice lines, we demonstrated that Cas9-ng had much higher editing efficiency than Cas9-ngv1 and xCas9 at ng PAM sites. To expand the base-editing scope, we developed an efficient C to T base-editing system by making fusion of Cas9-ng nickase (D10A version), PmCDAI, and UGI. Taken together, our work benchmarked xCas9 as a high-fidelity nuclease for targeting canonical ngG PAMs and Cas9-ng as a preferred variant for targeting relaxed PAMs for plant genome editing.

关键词： xCas9 Cas9-ng high fidelity non-canonical PAM base editing

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内镜检查：早期喉部和下咽部癌的NBI，AFI，和SMR诊断技术

内镜检查：早期喉部和下咽部癌的NBI，AFI，和SMR诊断技术

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中华医学会第十次全国耳鼻咽喉-头颈外科学术会议

作者： Hing-sang Chan Philip W.Y.Chiu Alexander Vlantis Raymond K.Y.Tsang C Andrew van Hasselt siu-Kwan ng Michael C.F.Tong Department of Otorhinolaryngology Head and Neck Surgerythe Chinese University of Hong Kong Department of Surgery The Chinese University of Hong KongHong Kong SAR

前言大部分头颈部鳞状上皮细胞癌(HNSCc)表现为处于进展期，大范围的手术切除会造成严重的功能障碍和美容上的畸形，总存活数常让人失望。除了局部病变，同期或不同期SCC(短程化疗，小细胞癌)的发生对病死率产生显著影响。

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铜表面微蚀处理新技术

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电镀与精饰 2008年第4期30卷 4-7页

作者：郑博江建平寇文鹏吴荣生 Joe Abys simon WAng 乐思化学新加坡638382 广州确信乐思化学贸易有限公司广东深圳518055 Ehthone Inc. 美国CT 06516

采用过硫酸盐和几种特别添加剂配制成的微蚀剂,不仅具有传统微蚀剂的优点,而且拥有其独特的优点:微蚀处理的铜表面比传统微蚀剂处理的光亮;微蚀处理后暴露在空气中10 min,铜表面也不变色;微蚀处理后的元件没有原电池腐蚀现象发生;其工... 详细信息

采用过硫酸盐和几种特别添加剂配制成的微蚀剂,不仅具有传统微蚀剂的优点,而且拥有其独特的优点:微蚀处理的铜表面比传统微蚀剂处理的光亮;微蚀处理后暴露在空气中10 min,铜表面也不变色;微蚀处理后的元件没有原电池腐蚀现象发生;其工作溶液比传统微蚀剂稳定;微蚀速率容易控制以满足不同的微蚀要求;微蚀处理不影响精密线路板和元件的尺寸。

关键词：新微蚀剂微蚀速率抛光原电池腐蚀

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Role of interleukin-1 and its antagonism of hepatic stellate cell proliferation and liver fibrosis in the Abcb4^(-/-) mouse model

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World Journal of Hepatology 2016年第8期8卷 401-410页

作者： Florian P Reiter Ralf Wimmer Lena Wottke Renate Artmann Jutta M Nagel manuel O Carranza Doris Mayr Christian Rust Peter Fickert Michael Trauner Alexander L Gerbes simon Hohenester Gerald U Denk Department of Medicine Ⅱ Liver Center Munich University of Munich Institute of Pathology University of Munich Department of Medicine Ⅰ Krankenhaus Barmherzige Brüder Research Unit for Experimental and Molecular Hepatology Division of Gastroenterology and Hepatology Department of Internal Medicine Hans Popper Laboratory of Molecular Hepatology Division of Gastroenterology and Hepatology Department of Internal Medicine Ⅲ Medical University of Vienna Transplant Center Munich of the University of Munich

AIM: To study the interleukin-1(IL-1) pathway as a therapeutic target for liver fibrosis in vitro and in vivo using the ATP-binding cassette transporter b4^(-/-)(Abcb4^(-/-)) mouse ***: Female and male Abcb4^(-/-) mice from 6 to 13 mo of age were analysed for the degree of cholestasis(liver serum tests), extent of liver fibrosis(hydroxyproline content and Sirius red staining) and tissue-specific activation of signalling pathways such as the IL-1 pathway [quantitative polymerase chain reaction(q PCR)]. For in vivo experiments, murine hepatic stellate cells(HSCs) were isolated via pronasecollagenase perfusion followed by density gradient centrifugation using female mice. Murine HSCs were stimulated with up to 1 ng/m L IL-1β with or without 2.5 μg/m L Anakinra, an IL-1 receptor antagonist, respectively. The proliferation of murine HSCs was assessed via the Brd U assay. The toxicity of Anakinra was evaluated via the fluorescein diacetate hydrolysis(FDH) assay. In vivo 8-wk-old Abcb4^(-/-) mice with an already fully established hepatic phenotype were treated with Anakinra(1 mg/kg body-weight daily intraperitoneally) or vehicle and liver injury and liver fibrosis were evaluated via serum tests, q PCR, hydroxyproline content and Sirius red staining. RESULTS: Liver fibrosis was less pronounced in males than in female Abcb4^(-/-) animals as defined by a lower hydroxyproline content(274 ± 64 μg/g vs 436 ± 80 μg/g liver, respectively; n = 13-15; P mannWhitney U-test) and lower m RNA expression of the profibrogenic tissue inhibitor of metalloproteinase-1(TIMP)(1 ± 0.41 vs 0.66 ± 0.33 fold, respectively; n = 13-15; P mann-Whitney U-test). Reduced liver fibrosis was associated with significantly lower levels of F4/80 m RNA expression(1 ± 0.28 vs 0.71 ± 0.41 fold, respectively; n = 12-15; P mann-Whitney U-test) and significantly lower IL-1β m RNA expression levels(1 ± 0.38 vs 0.44 ± 0.26 fold, respectively; n = 13-15; P mann-Whitney U-te

关键词： Cholestasis Primary sclerosing cholangitis The ATP-binding cassette transporter b4 Liver fibrosis Interleukin-1

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Endoscopic submucosal dissection vs laparoscopic colorectal resection for early colorectal epithelial neoplasia

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World Journal of Gastrointestinal Endoscopy 2015年第17期7卷 1243-1249页

作者： Sophie SF Hon simon SM ng Tiffany CL Wong Philip WY Chiu Tony WC Mak WW Leung Janet FY Lee Department of Surgery Prince of Wales HospitalThe Chinese University of Hong Kong

AIM: To compare the short term outcome of endoscopic submucosal dissection(ESD) with that of laparoscopic colorectal resection(LC) for the treatment of early colorectal epithelial neoplasms that are not amenable to conventional endoscopic removal. METHODS: This was a retrospective cohort study. The clinical data of all consecutive patients who underwent ESD for endoscopically assessed benign lesions that were larger than 2 cm in diameter from 2009 to 2013 were collected. These patients were compared with a cohort of controls who underwent LC from 2005 to 2013. Lesions that were proven to be malignant by initial endoscopic biopsies were excluded. Mid and lower rectal lesions were not included because total mesorectal excision, which bears a more complicated postoperative course, is not indicated for lesions without histological proof of malignancy. Both ESD and LC were performed by the same surgical unit with a standardized technique. The patients were managed according to a standard protocol, and they were closely monitored for complications after the procedures. All hospital records were reviewed, and the following data were compared between the ESD and LC groups: patient demographics, size and location of the lesions, procedure time, shortterm clinical outcomes and pathology results. RESULTS: From 2005 to 2013, 65 patients who underwent ESD and 55 patients who underwent LC were included in this study. The two groups were similar in terms of sex(P = 0.41) and American Society of Anesthesiologist class(P = 0.58), although patients in the ESD group were slightly older(68.6 ± 9.4 vs 64.6 ± 9.9, P = 0.03). ESD could be accomplished with a shorter procedure time(113 ± 66 min vs 153 ± 43 min, P < 0.01) for lesions of comparable size(3.0 ± 1.2 cm vs 3.4 ± 1.4 cm, P = 0.22) and location(colon/rectum:59/6 vs colon/rectum: 52/3, P = 0.43). ESD appeared to be associated with a lower short-term complication rate, but the difference did not reach statistical significance(10.8%

关键词： Early colorectal neoplasia Laparoscopic colectomy

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Frequency of Hypovitaminosis D in 6 - 59 Month Children with Severe Malaria in the Pediatrics Unit of the Teaching Hospital of Parakou (CHUD/BA) in Benin in 2016

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Open Journal of Pediatrics 2018年第1期8卷 66-73页

作者： Alphonse Noudamadjo Julien Didier Adédémy Joseph Agossou Gratien Godonou Sagbo Falilatou Agbeille Gérard Kpanidja Jacques Assoklé simon Akpona Faculty of Medicine University of Parakou CHUD/BA Parakou Benin Faculty of Health Sciences of Cotonou Cotonou Benin

Introduction: Vitamin D’s action outside of bone, especially on immunity, is widely reported in the international scientific literature over the last years. Objective: Calculate the frequency of hypovitaminosis D in children aged 6 to 59 months suffering from severe malaria in the CHUD-P pediatric unit in 2016. Setting and Methods: This research work is a cross-sectional study with descriptive and analytical purposes. Data gathering was prospective. The study involved children aged 6 to 59 months hospitalized for severe malaria in the CHUD-P pediatric unit. The said children were HIV-uninfected, eutrophic and had not received vitamin D supplementation during the last 6 months. Vitamin D dose was measured using the High Performance Liquid Chromatography (HPLC) technique. Results: A total of 80 subjects were involved in the survey. Mean age was 26.08 months, sex ratio was 0.8 and average weight was 10.80 kg. Hypovitaminosis D frequency was 83.8% (67 cases out of 80 children investigated during the survey) with an average plasma concentration of vitamin D estimated at 21.57 ng/ml ± 7.34 with two extremes (11.24 - 42.32) ng/ml. The minimum parasitaemia was 202 P/μl and the maximum was 580,000 P/μl. Conclusion: Hypovitaminosis D is common in children suffering from severe malaria;this result suggests conducting a large-scale community-based study to decide on vitamin D inclusion in national supplementation policies and severe malaria management.

关键词： Hypovitaminosis D Severe Malaria Children Benin

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Serum Levels of Prostate Specific Antigen Parameters among Male Patients with Lepromatous Leprosy in Edo State, Nigeria

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Health 2021年第9期13卷 920-933页

作者： Babatunde Ishola Gabriel Adejumo David Osebhor Uchechukwu Dimkpa Usman Itakure Abdulkadir Oladimeji Nasiru Abdulrahman Uzor simon Kelechi Ncheta Nkwazema Noreen Ebelechukwu Agbapuonwu Grace Umahi-Ottah Medical Laboratory Science Department University of Benin Benin City Nigeria Physiology Department Nnewi Campus Nnamdi Azikiwe University Awka Nigeria Department of Medical Laboratory Science College of Health Sciences and Technology Markafi Kaduna State Nigeria Department of Medical Laboratory Science College of Health Technology Offa Kwara State Department of Applied Science Faculty of Health and Applied Sciences University of West of England Bristol UK Medical Laboratory Science Department National Ear Care Centre Kaduna Kaduna State Nigeria Department of Nursing Nnamdi Azikwe University Akwa Nigeria Department of Physiology Ebonyi State University Abakaliki Nigeria

Aim: This study is aimed at determining the total PSA (tPSA), free PSA (fPSA) concentrations and free to total PSA ratio (f/tPSA) and their utility in prostate cancer (PCa) risk stratification among male lepromatous l... 详细信息

ng>Aim: ng>This study is aimed at determining the total PSA (tPSA), free PSA (fPSA) concentrations and free to total PSA ratio (f/tPSA) and their utility in prostate cancer (PCa) risk stratification among male lepromatous leprosy patients in Edo state, Nigeria. ng>Methods:ng> Forty participants and thirty controls participated in this study. A structured questionnaire was administered to each participant to elicit details such as their personal data, age, sex, residence, medications, alcohol consumption, smoking habit, length of time of diagnosis, sexual activities, as well as the history of any other underlying diseases. Blood samples were collected from the participants. Total and free PSA concentrations in the samples were measured using ELISA method. ng>Results:ng> Significantly (p = 0.042) higher serum fPSA among the leprosy patients (0.22 ± 0.12 ng/ml) compared with the healthy controls (0.17 ± 0.09 ng/ml). There were no significant (p = 0.055) difference in the mean tPSA between the leprosy patients (2.69 ± 2.25 ng/ml) compared with the control (1.95 ± 0.69 ng/ml). Similarly, no significant (p = 0.548) difference was observed in PSA ratio between the control (0.08 ± 0.04) and the leprosy patients (0.09 ± 0.03). ng>Conclusion:ng> Our study indicated no significant differences in tPSA and f/tPSA between the leprosy patients and their healthy control. Elevated tPSA as well as f/tPSA levels that fell within the high risk zones of PCa were more prevalent among patients aged ≥ 70 years, not married, who rarely had sexual intercourse and those with longer disease duration.

关键词： Serum Prostate Cancer Total PSA Free PSA Male Leprosy

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间皮瘤的根治手术仍处于流行高峰,需要进一步探索处理方法

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英国医学杂志中文版 2004年第3期7卷 138-139页

作者： Tom Treasure David Waller simon Swift Julian Peto 王燕 professor Cardiothoracic Unit Guy's Hospital London SEl 9RT consultant thoracic surgeon Glenfield Hospital Leicester LE3 9QP research registrar Thoracic Surgery Guy's Hospital London SEl 9RT professor of epidemiology Section of Epidemiology Institute of Cancer Research Sutton Surrey SM2 5NG

仍处于流行高峰。

关键词：根治术外科治疗间皮瘤石棉

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Reproductive Hormone Profiles and Their Relationships with Sexual Function in Patients with Prostate Cancer after Rapid Prostatectomy in Benin City

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Advances in Sexual Medicine 2020年第4期10卷 119-130页

作者： Babatunde Ishola Gabriel Adejumo Omolei Lilian Williams Efosa Bolaji Odigie Ifeoma Gloria Unachukwu Oladimeji Nasiru Abdulrahman Uchechukwu Dimkpa simon Uzor Oyarazi Margaret Adebowale Ojo Moses Oke Department of Medical Laboratory Science University of Benin Benin Nigeria Department of Medical Laboratory Science College of Health Technology Ilorin Nigeria Department of Human Physiology Nnamdi Azikwe University Nnewi Campus Awka Nigeria Department of Applied Science Faculty of Health and Applied Sciences University of West of England Bristol UK Department of Virology/Immunology Federal School of Medical Laboratory Science Jos Nigeria Department of Medical Laboratory Science College of Health Technology Akure Nigeria

There are limited studies examining the relationship between serum reproductive hormone levels and sexual functions among prostate cancer (PCa) patients after rapid prostatectomy (RP). Aim: The present study aimed at evaluating the correlation between post-operative serum levels of some male reproductive hormones of PCa patients and their sexual functions in the months following RP. A total of 20 male patients with clinically localized PCa who had undergone RP within the last 12 months prior to commencement of the study, were invited to participate. The postoperative serum levels of their luteinizing hormone (LH), follicle stimulating hormone (FSH) and total testosterone (Te) were measured. They also completed a structured health and lifestyle questionnaire to obtain information on their demographic characteristics and detailed medical history. Our data indicated normal mean levels of post-operative Te (4.34 ± 4.38 ng/ml) in the patients following RP, while the mean levels of FSH (39.24 ± 34.12 miu/ml) and LH (21.67 ± 25.73 miu/ml) were on the increase and far above normal ranges for healthy men. Data indicated a significant positive correlation between Te and frequency of sexual intercourse (r = 508;p < 0.05), libido (r = 0.429;p <

关键词： Reproductive Hormone Sexual Function Prostate Cancer Rapid Prostatectomy

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一种新型的高清晰度动态方法对年龄相关性黄斑变性视野中暗点的定位

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世界核心医学期刊文摘（眼科学分册） 2005年第2期1卷 45-46页

作者： Zur D. simon G.J.B. Loewenstein A. 肖颖 Jules Stein Eye I nstitute 100 Stein Plaza LosAngeles CA 90095 United States Dr.

BACKGROUND AND OBJECTIVE: To examine a new highresolution kinetic mapping meth od for scotoma in age-related macular degeneration. PATIENTS AND METHODS: A com puter based program for kinetic visual field mapping was tested in 10 healthy su bjects and 14 patients with age-related macular degeneration and fixed preferre d retinal locus. The stimulus was presented using a back projector on a screen l ocated 40 cm from the subject. The findings were then compared with static resul ts. RESULTS:Control group mapping revealed good congruency with the anatomic bli nd spot. Mapping of the 14 patients with age-related macular degeneration was r apid and revealed good accuracy. The average deviation of the mapping border from the a natomic scotoma border was no more than 3.1%of the scotoma radius. Static mappi ng of 7 of the patients with age-related macular degeneration was longer and re vealed lower accuracy. CONCLUSIONS: The proposed method is more rapid, accurate, and consistent than static mapping. It allows accurate mapping of central scoto ma with suprathreshold stimulus, and may be used in the future for detecting the early stages of age-related macular degeneration using subthreshold stimulus.

关键词：动态方法黄斑病变年龄相关性生理盲点刺激信号性佳阈下刺激计算机程序动态法平均偏差

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