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Establishment of a Lateral Flow Dipstick Detection Method for Influenza A Virus Based on CRISPR/Cas12a System

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China CDC weekly 2024年第37期6卷 946-952,I0008页

作者： Xiaoyan Zhao Ximing Zheng Xiyong Yang Qi Guo Yi Zhang Jun Lou Department of Clinical laboratory Zhumadian Central HospitalZhumadian CityHenan ProvinceChina laboratory of Virology Beijing Key Laboratory of Etiology of Viral Diseases in ChildrenCapital Institute of PediatricsBeijingChina National Institute for viral Disease Control and Prevention Chinese Center for Disease Control and PreventionBeijingChina

Objective:This study aimed to develop a rapid,visual PCR-CRISPR/Cas12-LFD method for detecting influenza A by utilizing the conserved region of the matrix protein ***:We crafted universal degradation primers and clustered regularly interspaced short palindromic repeats RNA(CRISPR RNA,crRNA)targeting the conserved matrix protein gene of the influenza virus(IFV),integrated with lateral flow dipstick(LFD)*** new PCR-CRISPR/Cas12-LFD approach was designed to determine its sensitivity and specificity through the analysis of various clinical samples collected in ***:The developed nucleic acid assay for influenza A viruses(IAV)demonstrated a sensitivity of 10 copies/μL without cross-reactivity with other respiratory *** of 82 clinical samples showed high concordance with results from fluorescent Polymerase Chain Reaction(PCR),achieving a kappa value of ***:A highly sensitive and specific PCRCRISPR/Cas12-LFD method has been successfully established for the detection of influenza A,offering a robust tool for its diagnosis and aiding in the prevention and control of this virus.

关键词： prevention diagnosis specificity

Mechanical regulation of lipid and sugar absorption by Piezo1 in enterocytes

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Acta Pharmaceutica Sinica B 2024年第8期14卷 3576-3590页

作者： Tian Tao Qing Shu Yawen Zhao Wenying Guo Jinting Wang Yuhao Shi Shiqi Jia Hening Zhai Hui Chen Cunchuan Wang Geyang Xu Department of Physiology School of MedicineJinan UniversityGuangzhou 510632China Department of Metabolic and Bariatric Surgery the First Affiliated Hospital of Jinan UniversityGuangzhou 510630China Endoscopy Center the First Affiliated Hospital of Jinan UniversityGuangzhou 510630China Biotherapy Center Cell-gene Therapy Translational Medicine Research Centerthe Third Affiliated HospitalSun Yat-sen UniversityGuangzhou 510630China Key laboratory of viral Pathogenesis&Infection Prevention and Control(Jinan University) Ministry of EducationGuangzhou 510632China

Obesity is primarily caused by excessive intake as well as absorption of sugar and *** surge in distention pressure and intestinal motility accelerates the absorption of *** response of intestinal epithelial cells to mechanical stimulation is not fully ***1,a mechanosensitive ion channel,is widely expressed throughout the digestive ***,its function in intestinal nutrient absorption is not yet *** our study,excessive lipid deposition was observed in the duodenum of obese patients,while duodenal Piezo1-CaMKK2-AMPKa was decreased when compared to normal-weight *** high-fat diet condition,the Piezo1iKO mice exhibited abnormally elevated sugar and lipid absorption as well as severe lipid deposition in the duodenum and *** phenotypes were mainly caused by the inhibition of duodenal CaMKK2-AMPKa and the upregulation of SGLT1 and *** contrast,Yoda1,a Piezo1 agonist,was found to reduce intestinal lipid absorption in diet induced obese *** of Piezo1,stretch and Yoda1 inhibited lipid accumulation and the expression of DGAT2 and SGLT1,whereas knockdown of Piezo1 stimulated lipid accumulation and DGAT2 in Caco-2 *** study reveals a previously unexplored mechanical regulation of nutrient absorption in intestinal epithelial cells,which may shed new light on the therapy of obesity.

关键词： Piezo1 DGAT2 SGLT1 Lipid metabolism Obesity Mechanical sensing Intestinal lipid absorption Intestinal glucose absorption

A Non-canonical Excitatory PV RGC–PV SC Visual Pathway for Mediating the Looming-evoked Innate Defensive Response

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Neuroscience Bulletin 2024年第3期40卷 310-324页

作者： Man Yuan Sen Jin Gao Tan Siyuan Song Yizong Liu Huadong Wang Yin Shen Eye Center Renmin Hospital of Wuhan UniversityWuhan UniversityWuhan430060China The Brain Cognition and Brain Disease Institute Shenzhen Key Laboratory of Viral Vectors for BiomedicineShenzhen Institute of Advanced TechnologyChinese Academy of Sciences(CAS)Shenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research InstitutionsNational Medical Products Administration Key Laboratory for Research and Evaluation of Viral Vector Technology in Cell and Gene Therapy Medicinal ProductsShenzhen Key Laboratory of Quality Control Technology for Virus-Based TherapeuticsGuangdong Provincial Medical Products AdministrationShenzhen518055China Jan and Dan Duncan Neurological Research Institute Baylor College of MedicineHouston77030USA State Key laboratory of Magnetic Resonance and Atomic and Molecular Physics Key Laboratory of Magnetic Resonance in Biological SystemsWuhan Center for Magnetic ResonanceInnovation Academy for Precision Measurement Science and TechnologyCASWuhan430071China Frontier Science Center of Immunology and Metabolism Medical Research InstituteWuhan UniversityWuhan430071China

Parvalbumin-positive retinal ganglion cells(PV+RGCs)are an essential subset of RGCs found in various ***,their role in transmitting visual information remains ***,we characterized PV+RGCs in the retina and explored the functions of the PV+RGC-mediated visual *** applying multiple viral tracing strategies,we investigated the downstream of PV+RGCs across the whole ***,we found that the PV+RGCs provided direct monosynaptic input to PV+excitatory neurons in the superficial layers of the superior colliculus(SC).Ablation or suppression of SC-projecting PV+RGCs abolished or severely impaired the flight response to looming visual stimuli in mice without affecting visual ***,using transcriptome expression profiling of individual cells and immunofluorescence colocalization for RGCs,we found that PV+RGCs are predominant glutamatergic ***,our findings indicate the critical role of PV+RGCs in an innate defensive response and suggest a non-canonical subcortical visual pathway from excitatory PV+RGCs to PV+SC neurons that regulates looming visual *** results provide a potential target for intervening and treating diseases related to this circuit,such as schizophrenia and autism.

关键词： Parvalbumin-positive retinal ganglion cell Innate fear Superior colliculus Excitatory-excitatory neuronal connection Looming-evoked defensive response Subcortical pathway

Development of an Integrated Disposable Device for SARSCoV-2 Nucleic Acid Extraction and Detection

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Biomedical and Environmental Sciences 2024年第6期37卷 639-646页

作者： Jing Ma Yanzhe Hao Meiling Hou Xiaoshan Zhang Jingduan Liu Haodi Meng Jiangbo Chang Xuejun Ma Jihua Liu Qingjie Ying Xianhua Wang Hongxia Li Yuxi Cao Xiaoguang Zhang National Key laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China Beijing Runbio Biotechnology Development Co. LtdBeijing 100106China Beijing Beta Technology Co. LtdBeijing 101318China Jiangsu Qitian gene Biotechnology Co. LtdWuxi 214135JiangsuChina

Objective To develop a highly sensitive and rapid nucleic acid detection method for the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Methods We designed,developed,and manufactured an integrated disposable device for SARS-CoV-2 nucleic acid extraction and *** precision of the liquid transfer and temperature control was tested.A comparison between our device and a commercial kit for SARS-Cov-2 nucleic acid extraction was performed using real-time fluorescence reverse transcription polymerase chain reaction(RT-PCR).The entire process,from SARS-CoV-2 nucleic acid extraction to amplification,was *** The precision of the syringe transfer volume was 19.2±1.9μL(set value was 20),32.2±1.6(set value was 30),and 57.2±3.5(set value was 60).Temperature control in the amplification tube was measured at 60.0±0.0℃(set value was 60)and 95.1±0.2℃(set value was 95)***-Cov-2 nucleic acid extraction yield through the device was 7.10×10^(6) copies/mL,while a commercial kit yielded 2.98×10^(6) copies/*** mean time to complete the entire assay,from SARS-CoV-2 nucleic acid extraction to amplification detection,was 36 min and 45 *** detection limit for SARS-CoV-2 nucleic acid was 250 copies/*** The integrated disposable devices may be used for SARS-CoV-2 Point-of-Care test(POCT).

关键词： An integrated disposable device SARS-Cov-2 Nucleic acid detection

Engineered AAV13 variants with enhanced transduction and confined spread

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Zoological Research 2024年第4期45卷 781-790页

作者： Neng-Song Luo Yu-Xiang Cai Zeng-Peng Han Xiao-Kai Sui Wen-Jia Yuan Zi-Lian Zhang Hao-Dong Guo Jie Wang Kun-Zhang Lin Fu-Qiang Xu Wuhan National laboratory for Optoelectronics Huazhong University of Science and TechnologyWuhanHubei 430074China Shenzhen Key laboratory of viral Vectors for Biomedicine Shenzhen-Hong Kong Institute of Brain ScienceShenzhen Institute of Advanced TechnologyChinese Academy of SciencesShenzhenGuangdong 518055China Key laboratory of Magnetic Resonance in Biological Systems State Key Laboratory of Magnetic Resonance and Atomic and Molecular PhysicsNational Center for Magnetic Resonance in WuhanInnovation Academy for Precision Measurement Science and TechnologyChinese Academy of SciencesWuhanHubei 430071China Key laboratory of Quality Control Technology for Virus-Based Therapeutics Guangdong Provincial Medical Products AdministrationNMPA Key Laboratory for Research and Evaluation of Viral Vector Technology in Cell and Gene Therapy Medicinal ProductsBrain Cognition and Brain Disease InstituteShenzhen Institute of Advanced TechnologyChinese Academy of SciencesShenzhenGuangdong 518055China University of Chinese Academy of Sciences Beijing 100049China Zhongnan Hospital Wuhan UniversityWuhanHubei 430072China Brain Case Biotechnology Co. Ltd.ShenzhenGuangdong 518107China College of Life Sciences Wuhan UniversityWuhanHubei 430072China Center for Excellence in Brain Science and Intelligence Technology Chinese Academy of SciencesShanghai 200031China

Precise targeting of specific regions within the central nervous system(CNS)is crucial for both scientific research and gene therapy in the context of brain ***-associated virus 13(AAV13)is known for its restricted diffusion range within the CNS,making it an ideal choice for precise labeling and administration within small brain ***,AAV13 mediates relatively low expression of target ***,we introduced specifically engineered modifications to the AAV13 capsid protein to enhance its transduction *** first constructed AAV13-YF by mutating tyrosine to phenylalanine on the surface of the AAV13 *** then inserted the 7m8 peptide,known to enhance cell transduction,into positions 587/588 and 585/586 of the AAV13 capsid,resulting in two distinct variants named AAV13-587-7m8 and AAV13-585-7m8,*** found that AAV13-YF exhibited superior in vitro infectivity in HEK293T cells compared to AAV13,while AAV13-587-7m8 and AAV13-585-7m8 showed enhanced CNS infection capabilities in C57BL/6 mice,with AAV13-587-7m8 infection retaining a limited spread *** modified AAV13 variants hold promising potential for applications in gene therapy and neuroscience research.

关键词： Adeno-associated virus 13 AAV13-YF AAV13-587-7m8 AAV13-585-7m8 transduction efficiency

Seasonal and genetic Characteristics of Human Metapneumovirus Circulating—Henan Province,China,2017-2023

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China CDC weekly 2024年第20期6卷 450-456,I0001页

作者： Zhibo Xie Zhen Zhu Jin Xu Naiying Mao Aili Cui Wenhui Wang Yage Wang Zhibo Zhang Baicheng Xia Haoran Wang Zhen Sun Yan Zhang National Key laboratory of Intelligent Tracking and Forecasting for Infectious Diseases NHC Key Laboratory of Medical Virology and Viral DiseaseNational Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijingChina Henan Provincial Center for Disease Control and Prevention Luohe CityHenan ProvinceChina Luohe Center for Disease Control and Prevention Luohe CityHenan ProvinceChina Luohe Central Hospital Luohe CityHenan ProvinceChina

Introduction:This study examines the seasonal and genetic characteristics of human metapneumovirus(HMPV)in Henan from 2017 to ***:Samples from patients with acute respiratory infection(ARI)testing positive for HMPV were subjected to real-time reverse transcription polymerase chain reaction The G gene was amplified and sequenced from these samples for epidemiological and phylogenetic ***:We enrolled 2,707 ARI patients from October 2017 to March 2023,finding an HMPV positivity rate of 6.17%(167/2,707).Children under five exhibited the highest infection rate at 7.78%(138/1,774).The 2018 and 2019 HMPV outbreaks predominantly occurred in spring(March to May),with peak positivity rates of 31.11%in May 2018 and 19.57%in May 2019.A notable increase occurred in November 2020,when positivity reached a historic high of 42.11%,continuing until January *** February 2021 through March 2023,no significant seasonal peaks were observed,with rates ranging from 0%to 8.70%.Out of 81 G gene sequences analyzed,46.91%(38/81)were identified as subtype A(A2c:45.67%,37/81;A2b:1.23%,1/81)and 53.09%(43/81)as subtype B(B1:9.88%,8/81;B2:43.21%,35/81).Notably,an AAABBA switch pattern was observed in HMPV *** dominant strains were A2c111nt-dup in subtype A and B2 in subtype ***:Six years of surveillance in Henan Province has detailed the seasonal and genetic dynamics of HMPV,contributing valuable insights for the control and prevention of HMPV infections in *** findings support the development of targeted HMPV vaccines and immunization strategies.

关键词： prevention finding seasonal

Exploiting viral vectors to deliver genome editing reagents in plants

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aBIOTECH 2024年第2期5卷 247-261页

作者： Yilin Shen Tao Ye Zihan Li Torotwa Herman Kimutai Hao Song Xiaoou Dong Jianmin Wan State Key laboratory of Crop genetics and Germplasm Enhancement and Utilization Jiangsu Collaborative Innovation Centre for Modern Crop ProductionJiangsu Engineering Research Center for Plant Genome EditingNanjing Agricultural UniversityNanjing 210095China Zhongshan Biological Breeding laboratory Nanjing 210014China Hainan Seed Industry laboratory Sanya 572025China State Key laboratory of Crop gene Resources and Breeding Institute of Crop SciencesChinese Academy of Agricultural SciencesBeijing 100081China

Genome editing holds great promise for the molecular breeding of plants,yet its application is hindered by the shortage of simple and effective means of delivering genome editing reagents into *** plant transformation-based methods for delivery of genome editing reagents into plants often involve prolonged tissue culture,a labor-intensive and technically challenging process for many elite crop *** this review,we describe various virus-based methods that have been employed to deliver genome editing reagents,including components of the CRISPR/Cas machinery and donor DNA for precision editing in *** update the progress in these methods with recent successful examples of genome editing achieved through virus-based delivery in different plant species,highlight the advantages and limitations of these delivery approaches,and discuss the remaining challenges.

关键词： Plant genome engineering Genome editing CRISPR/Cas Virus-based delivery

To Analyze the Sensitivity of RT-PCR Assays Employing S gene Target Failure with Whole Genome Sequencing Data during Third Wave by SARS-CoV-2 Omicron Variant

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Advances in Microbiology 2024年第5期14卷 247-255页

作者： Pooja Patel Yogita Mistry Monika Patel Summaiya Mullan viral Research and Diagnostic laboratory Department of Microbiology Government Medical college Surat India

Introduction: Omicron is a highly divergent variant of concern (VOCs) of a severe acute respiratory syndrome SARS-CoV-2. It carries a high number of mutations in its spike protein hence;it is more transmissible in the community by immune evasion mechanisms. Due to mutation within S gene, most Omicron variants have reported S gene target failure (SGTF) with some commercially available PCR kits. Such diagnostic features can be used as markers to screen Omicron. However, Whole Genome Sequencing (WGS) is the only gold standard approach to confirm novel microorganisms at genetically level as similar mutations can also be found in other variants that are circulating at low frequencies worldwide. This Retrospective study is aimed to assess RT-PCR sensitivity in the detection of S gene target failure in comparison with whole genome sequencing to detect variants of Omicron. Methods: We have analysed retrospective data of SARS-CoV-2 positive RT-PCR samples for S gene target failure (SGTF) with TaqPath COVID-19 RT-PCR Combo Kit (ThermoFisher) and combined with sequencing technologies to study the emerged pattern of SARS-CoV-2 variants during third wave at the tertiary care centre, Surat. Results: From the first day of December 2021 till the end of February 2022, a total of 321,803 diagnostic RT-PCR tests for SARS-CoV-2 were performed, of which 20,566 positive cases were reported at our tertiary care centre with an average cumulative positivity of 6.39% over a period of three months. In the month of December 21 samples characterized by the SGTF (70/129) were suggestive of being infected by the Omicron variant and identified as Omicron (B.1.1.529 lineage) when sequence. In the month of January, we analysed a subset of samples (n = 618) with SGTF (24%) and without SGTF (76%) with Ct values Conclusions: During the COVID-19 pandemic, it took almost more than 15 days to diagnose infection and identify pathogen by sequencing technology. In contrast to that molecular assay provided qu

关键词： SARS-CoV-2 S gene Target Failure Whole Genome Sequencing Omicron

PREPARATION OF gene-viral THERAPEUTIC SYSTEM CNHK200-HA AND ITS ANTITUMOR ACTIVITY ON LUNG CANCER

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Chinese Journal of Cancer Research 2006年第1期18卷 1-7页

作者：王伟国马炬明苏长青胡慧珍钱其军 Department of Internal Medicine Hangzhou 117 Hospital Hangzhou 310004 Department of viral and gene therapy Eastern Hepatobiliary Surgery Hospital Second Military Medical University Shanghai 200438

Objective： To develop a novel adenoviral vector system, which combines the advantages of the antiangiogenic gene therapy and virus therapy, and to investigate its antitumor activity on lung cancer. Methods： A new kind of viral vector CNHK200, in which the Elb55kDa gene was deleted and the whole Ela gene was preserved, was constructed. Human angiostatin gene Kringle 1-5 （hA） was amplified and inserted into the genome of the replicative virus CNHK200, generating CNHK200-hA. The expression of hA and its effect on lung cancer cell growth in vitro and in vivo were studied. Results： The novel vector system CNHK200-hA, just like the replicative virus ONYX-015, replicated in p53-deficient tumor cells but not in normal cells, and thus specifically killed tumor cells. In in vitro experiment, both CNHK200-hA and the non-replicative virus Ad-hA could kill tumor cells but the latter needed 100 times more MOI to achieve the same level of cell killing. In in vivo experiment, the therapeutic effect of CNHK200-hA on human lung cancer A549 xenografts in nude mice was significantly better than that of Ad-hA or that of ONYX-015. Conclusion： CNHK200-hA, which carries the angiostatin gene, has the advantages of specific tumor targeting, high expression of transgene in tumor cells and potent antitumor activity.

关键词： Adenovirus/replicative gene therapy Antiangiogenesis Lung cancer