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| 1 | Inhibiting effect of antisense oligonucleotides phosphorthioate on gene expression of TIMP-1 in rat liver fibrosis显示文摘AIM To observe the inhibition of antisenseoligonucleotides (asON) phosphorthioate to thetissue inhibitors metalloproteinase-1 (TIMP-1)gene and protein expression in the liver tissue ofimmunologically induced hepatic fibrosis rats.The possibility of reversing hepatic fibrosisthrough gene therapy was observed.METHODS Human serum albumin (HSA) wasused to attack rats, as hepatic fibrosis model, inwhich asONs were used to block the gene andprotein expressing TIMP-1. According to theanalysis of modulator, structure protein, codingseries of TIMP-1 genome, we designed fourdifferent asONs. These asONs were injected intothe hepatic fibrosis models through coccygealvein. The results was observed by RT-PCR formeasuring TIMP-1 mRNA expression,immunohistochemistry and in situ hybridizationfor collagen Ⅰ, Ⅲ, special staining of collagenfiber, and electron microscopic examination.RESULTS Hepatic fibrosis could last within 363days in our modified model. The expressinglevel of TIMP-1 was high during hepatic fibrosisprocess. It has been proved by theimmunohistochemical and the electronmicroscopic examination that the asONphosphorthioate of TIMP-1 could exactly expressin vivo. The effect of colchicine wasdemonstrated to inhibit the expressing level ofmRNA and the content of collagen Ⅰ, Ⅲ in theliver of experimental hepatic fibrosis rats.However, the electron microscopy research andthe pathologic grading of hepatic fibrosisshowed that there was no significant differencebetween the treatment group and the modelgroup (P>0.05).CONCLUSION The experimental rat model ofhepatic fibrosis is one of the preferable modelsto estimate the curative effect of anti-hepaticfibrosis drugs. The asON phosphorthioate ofTIMP-1 could block the gene and proteinexpression of TIMP-1 in the liver of experimentalhepatic fibrosis rats at the mRNA level. It ispossible to reverse hepatic fibrosis, and it isexpected to study a new drug of anti-hepaticfibrosis on the genetic level. Colchicine has verylimited therapeutic effect on hepatic fibrosis,furthermore, its toxicity and side effects areobvious. | Qing He Nie Yong Qian Cheng Yu Mei Xie Yong Xing Zhou Yi Zhan Cao The Center of Infectious Disease Diagnosis and Treatment of PLA,Tangdu Hospital,Forth Military Medical University,Xi’an 710038,Shaanxi Province,ChinaDr,Qing He Nie graduated from Qinghai Medical College as a doctor in 1983,got master degree at Beijing 302 Army Hospital in 1993,got doctor degree at the Third Military Medical University in 1998,engaged in postdoctoral research at the Fourth Military Medical University from 1998 to 2000,now an associate professor,specialized in clinical and experimental research of infectious diseases,had more than 90 papers published,coauthor of ten books,first author of one book. | 2001 | World Journal of Gastroenterology2001，7，3： | 73 |
| 2 | Polymorphisms in PPARD,PPARG and APM1 associated with four types of Traditional Chinese Medicine constitutions显示文摘Based on the theory of constitution of Traditional Chinese Medicine (TCM), the human population is divided into nine constitutions including one balanced constitution (Normality) and eight unbalanced constitutions (Yang-deficiency, Yin-deficiency, Phlegm-wetness, Qi-deficiency, Wetness-heat, Blood stasis, Depressed constitution, and Inherited special constitution). Different constitutions have specific metabolic features and different susceptibility to certain diseases. However, whether a genetic basis accounts for such constitution classifica-tion is yet to be determined. Here we performed a genetic study to assess the association between genetic variations of metabolic genes in-cluding PPARD, PPARG and APM1 and the constitutions. A total of 233 individuals of the Han population in China were classified into four groups, Normality, Yang-deficiency, Yin-deficiency and Phlegm-wetness with whom 23 single nucleotide polymorphisms (SNPs) in the three genes were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Biased distri-bution of PPARD rs2267669 and rs2076167, APM1 rs7627128 and rs1063539 in Yang-deficiency, PPARG Pro12Ala in Yin-deficiency and PPARD rs2076167, APM1 rs266729 and rs7627128 in Phlegm-wetness were observed. The frequencies of Haplotype13 (Hap13) of PPARG in Yin-deficiency, Hap25 of APM1 in Yang-deficiency and Hap2 of PPARD and Hap14 of PPARG in Phlegm-wetness, were significantly different from those in Normality, suggesting those might be group-associated haplotypes. These results suggested that single SNP and hap-lotypes of PPARD, PPARG and APM1 may underlie the genetic basis of the constitutions classified in TCM. | Yanrui Wu Yina Cun Jing Dong Jingru Shao Shengjun Luo Shengjie Nie Haijing Yu Bingrong Zheng Qi Wang Chunjie Xiao | 2010 | Journal of Genetics and Genomics2010，37，6： | 36 |
| 3 | 5-Hydroxymethylcytosine signatures in circulating cell-free DNA as diagnostic biomarkers for human cancers显示文摘象 5-methylcytosine (5mC ) 和 5-hydroxymethylcytosine (5hmC ) 那样的 DNA 修正是知道在哺乳动物影响全球基因表示的 epigenetic 标记。在人的染色体,与基因表示的靠近的关联和高化学的稳定性给他们的流行,这些 DNA epigenetic 标记能为癌症用作理想的 biomarkers 诊断。利用一种高度敏感、选择的化学标记技术,我们这里报导在传播没有房间的 DNA ( cfDNA )并且在从最近与 colorectal 诊断的 260 个病人的一个队收集的配对的肿瘤和邻近的纸巾的 genomic DNA ( gDNA )的 5hmC 的染色体宽的介绍,胃,胰腺,从 90 个健康个人的肝或甲状腺癌症和正常纸巾。5hmC 主要在与开的染色质和容许的 histone 修正重合的 transcriptionally 活跃的区域被散布。柔韧的联系癌症的 5hmC 签名在 cfDNA 被识别为特定的癌症类型是特征的。传播 cfDNA 的 5hmC 底 biomarkers colorectal 和胃的癌症是高度预兆的并且比常规 biomarkers 优异并且比得上从织物活体检视的 5hmC biomarkers。因此,这新策略能从血样品的分析为癌症的诊断和预后导致有效、最低限度地侵略的方法的发展。 | Wenshuai Li Xu Zhang Xingyu Lu Lei You Yanqun Song Zhongguang Luo Jun Zhang Ji Nie Wanwei Zheng Diannan Xu Yaping Wang Yuanqiang Dong Shulin Yu Jun Hong Jianping Shi Hankun Hao Fen Luo Luchun Hua Peng Wang Xiaoping Qian Fang Yuan Lianhuan Wei Ming Cui Taiping Zhang Quan Liao Menghua Dai Ziwen Liu Ge Chen Katherine Meckel Sarbani Adhikari Guifang Jia Marc B Bissonnette Xinxiang Zhang Yupei Zhao Wei Zhang Chuan He Jie Liu | 2017 | Cell Research2017，27，10： | 30 |
| 4 | Chinese Multidisciplinary Expert Consensus on the Diagnosis and Treatment of Hyperuricemia and Related Diseases显示文摘 | Zou, He-Jian Wu, Hu-Sheng Zhou, Jing-Guo Zeng, Xue-Jun Dai, Lie Wu, Hua-Xiang Zhu, Xiao-Xia Mei, Chang-Lin Hao, Chuan-Ming Chen, Nan Liu, Bi-Cheng Chen, Jiang-Hua Yang, Li Nie, Jing Yu, Chen Peng, Ai Yu, Sheng-Qiang Li, Lin Ge, Jun-Bo Huo, Yong Zhang, Shu-Yang Chen, Yun-Dai Dong, Yu-Gang Liang, Chun Dai, Yu-Xiang Gao, Xin Li, Chang-Gui Zhao, Jia-Jun Chen, Hai-Bing Cheng, Zhi-Feng Lin, Huan-Dong Guan, Yang-Tai Wang, Kai Luo, Ben-Yan Dai, Ruo-Lian Jiang, Quan Xue, Luan Liang, Chao-Chao Chen, Ming Fan, Song | 2017 | Chinese Medical Journal2017，，20： | 30 |
| 5 | Homology-mediated end joining-based targeted integration using CRISPR/Cas9显示文摘 | Yao, Xuan Wang, Xing Hu, Xinde Liu, Zhen Liu, Junlai Zhou, Haibo Shen, Xiaowen Wei, Yu Huang, Zijian Ying, Wenqin Wang, Yan Nie, Yan-Hong Zhang, Chen-Chen Li, Sanlan Cheng, Leping Wang, Qifang Wu, Yan Huang, Pengyu Sun, Qiang Shi, Linyu Yang, Hui | 2017 | Cell Research2017，27，6： | 28 |
| 6 | Acute inflammation stimulates a regenerative response in the neonatal mouse heart显示文摘 | Chunyong Han Yu Nie Hong Lian Rui Liu Feng He Huihui Huang Shengshou Hu | 2015 | Cell Research2015，25，10： | 26 |
| 7 | Simulation on dissolute and dust dispersion in comprehensive mechanized heading face with forced-exhaust ventilation显示文摘 | NIE Wen CHENG Wei-min HAN Li ZHOU Sheng-ju YU Yan-bin ZHAO Shan-shan | 2011 | Journal of Coal Science & Engineering(China)2011，17，3： | 21 |
| 8 | One-step generation of complete gene knockout mice and monkeys by CRISPR/Cas9-mediated gene editing with multiple sgRNAs显示文摘CRISPR/Cas9 系统是一个有效编辑基因的方法,但是编辑基因的动物的多数显示出 mosaicism,与编辑仅仅在房间的部分发生。这里,我们证明那单个基因或多重基因能被 Cas9 mRNA 和多重邻近的单个指南的 RNA 的 zygotic 注射完全在老鼠和猴子胚胎击倒(spaced 10-200 bp 分开) 指向仅仅一把单个钥匙每基因的 exon。个别地在 Y 染色体上跟随八基因的指向的删除的 F0 老鼠的 Phenotypic 分析在产生猛烈老鼠表明了这条途径的坚韧性。重要地,这条途径在高效率交付完全的基因大美人(Arntl 上的 100% 并且 91% 在 Prrt2 上) 在猴子胚胎。最后,我们能在一个单个步骤产生一只完全的 Prrt2 猛烈猴子,表明在很快建立编辑基因的猴子模型的这条途径的实用性。 | Erwei Zuo Yi-Jun Cai Kui Li Yu Wei Bang-An Wang Yidi Sun Zhen Liu Jiwei Liu Xinde Hu Wei Wei Xiaona Huo Linyu Shi Cheng Tang Dan Liang Yan Wang Yan-Hong Nie Chen-Chen Zhang Xuan Yao Xing Wang Changyang Zhou Wenqin Ying Qifang Wang Ren-Chao Chen Qi Shen Guo-Liang Xu Jinsong Li Qiang Sun Zhi-Qi Xiong Hui Yang | 2017 | Cell Research2017，27，7： | 21 |
| 9 | Antigen-presenting effects of effector memory Vγ9Vδ2 T cells in rheumatoid arthritis显示文摘 | Chaoying Hu Liu Qian Yi Miao Qiuyu Huang Ping Miao Ping Wang Qiwen Yu Hong Nie Jiying Zhang Dongyi He Rong Xu Xuehua Chen Bingya Liu Dongqing Zhang | 2012 | Cellular & Molecular Immunology2012，9，3： | 18 |
| 10 | Generation of a monkey with MECP2 mutations by TALEN-based gene targeting显示文摘Gene editing in model organisms has provided critical insights into brain development and diseases. Here, we report the generation of a cynomolgus monkey(Macaca fascicularis) carrying MECP2 mutations using transcription activator-like effector nucleases(TALENs)-mediated gene targeting. After injecting TALENs mRNA into monkey zygotes achieved by in vitro fertilization and embryo transplantation into surrogate monkeys, we obtained one male newborn monkey with an MECP2 deletion caused by frameshifting mutation in various tissues. The monkey carrying the MECP2 mutation failed to survive after birth, due to either the toxicity of TALENs or the critical requirement of MECP2 for neural development. The level of MeCP2 protein was essentially depleted in the monkey's brain. This study demonstrates the feasibility of introducing genetic mutations in non-human primates by site-specific gene-editing methods. | Zhen Liu Xue Zhou Ying Zhu Zhi-Fang Chen Bin Yu Yan Wang Chen-Chen Zhang Yan-Hong Nie Xiao Sang Yi-Jun Cai Yue-Fang Zhang Chen Zhang Wen-Hao Zhou Qiang Sun Zilong Qiu | 2014 | Neuroscience Bulletin2014，30，3： | 17 |
| 11 | Experimental study of bioartificial liver with cultured human liver cells显示文摘METHODSTheliversupportexperimentofEBLSSconsistingofaggregatesculturedhumanlivercels,holowfiberbioreactor,andcirculationunit... | WANG Ying Jie, LI Meng Dong, WANG Yu Ming, NIE Qing He and CHEN Guo Zheng | 1999 | World Journal of Gastroenterology1999，5，2： | 14 |
| 12 | Strontium ranelate reduces cartilage degeneration and subchondral bone remodeling in rat osteoarthritis model显示文摘 | De-gang YU Hui-feng DING Yuan-qing MAO Ming LIU Bo YU Xin ZHAO Xiao-qing WANG Yang LI Guang-wang LIU Shao-bo NIE Shen LIU Zhen-an ZHU | 2013 | Acta Pharmacologica Sinica2013，34，3： | 13 |
| 13 | Prediction of Primary Climate Variability Modes at the Beijing Climate Center显示文摘Climate variability modes, usually known as primary climate phenomena, are well recognized as the most important predictability sources in subseasonal–interannual climate prediction. This paper begins by reviewing the research and development carried out, and the recent progress made, at the Beijing Climate Center(BCC) in predicting some primary climate variability modes. These include the El Ni?o–Southern Oscillation(ENSO), Madden–Julian Oscillation(MJO), and Arctic Oscillation(AO), on global scales, as well as the sea surface temperature(SST) modes in the Indian Ocean and North Atlantic, western Pacific subtropical high(WPSH), and the East Asian winter and summer monsoons(EAWM and EASM, respectively), on regional scales. Based on its latest climate and statistical models, the BCC has established a climate phenomenon prediction system(CPPS) and completed a hindcast experiment for the period 1991–2014. The performance of the CPPS in predicting such climate variability modes is systematically evaluated. The results show that skillful predictions have been made for ENSO, MJO, the Indian Ocean basin mode, the WPSH, and partly for the EASM, whereas less skillful predictions were made for the Indian Ocean Dipole(IOD) and North Atlantic SST Tripole, and no clear skill at all for the AO, subtropical IOD, and EAWM. Improvements in the prediction of these climate variability modes with low skill need to be achieved by improving the BCC's climate models, developing physically based statistical models as well as correction methods for model predictions.Some of the monitoring/prediction products of the BCC-CPPS are also introduced in this paper. | Hong-Li REN Fei-Fei JIN Lianchun SONG Bo LU Ben TIAN Jinqing ZUO Ying LIU Jie WU Chongbo ZHAO Yu NIE Peiqun ZHANG Jin BA Yujie WU Jianghua WAN Yuping YAN Fang ZHOU | 2017 | Journal of Meteorological Research2017，31，1： | 13 |
| 14 | Plasma neutrophil-gelatinase-associated lipocalin and cystatin C could early diagnose contrast-induced acute kidney injury in patients with renal insufficiency undergoing an elective percutaneous coronary intervention显示文摘 | LIU Xiao-li WANG Zhi-jian YANG Qing YU Miao SHEN Hua NIE Bin HAN Hong-ya GAO Fei ZHOU Yu-jie | 2012 | Chinese Medical Journal2012，，6： | 12 |
| 15 | Expression and Comparative Analysis of Genes Encoding Outer Membrane Proteins LipL21,LipL32 and OmpL1 in Epidemic Leptospires显示文摘Leptospiral 外部膜蛋白质(OMP ) 高度在不同种类被保存,并且在新免疫保护和 serodiagnosis 策略的发展起一个必要作用。编码 LipL21, LipL32 和 OmpL1 的基因从螺旋体 interrogans serovar lai 种类 Lai 的完全的染色体顺序被克隆并且表示了试管内。顺序比较分析表明三基因高度在不同流行 leptospires 之中被保存,包括三主要流行种类螺旋体 interrogans,螺旋体 borgpetersenii 和螺旋体 weilii,在中国。Immunoblot 分析进一步被执行用重组体 OMP 的反时代细看 15 个流行螺旋体参考种类。两免疫污点试金和反向的抄写聚合酶链反应证明这三 OMP 保存地在病原的 L 被表示。interrogans 种类和另外的病原的 leptospires。另外,这些重组体 OMP 的使用作为在为细螺旋体病的 serodiagnosis 的连接酶的免疫吸着剂试金(ELISA ) 的抗原被评估。重组体 LipL32 和 OmpL1 蛋白质与重量的单位显示出 ELISA 反应的高度一感染 L 的 from 病人。interrogans 拉紧 Lai 和另外的病原的 leptospires。这些结果可以贡献宽广范围的疫苗和免疫的候选人的鉴定在进一步的研究的诊断抗原。 | Xiang-Yan ZHANG Yang YU Ping HE Yi-Xuan ZHANG Bao-Yu HU Yang YANG Yi-Xin NIE Xiu-Gao JIANG Guo-Ping ZHAO Xiao-Kui GUO | 2005 | Acta Biochimica et Biophysica Sinica2005，37，10： | 12 |
| 16 | Genetic lineage tracing identifies in situ Kit-expressing cardiomyocytes显示文摘 | Qiaozhen Liu Rui Yang Xiuzhen Huang Hui Zhang Lingjuan He Libo Zhang Xueying Tian Yu Nie Shengshou Hu Yan Yan Li Zhang Zengyong Qiao Qing-Dong Wang Kathy O Lui Bin Zhou | 2016 | Cell Research2016，26，1： | 12 |
| 17 | Experimental investigation on aero-heating of rudder shaft within laminar/turbulent hypersonic boundary layers显示文摘The aero-heating of the rudder shaft region of a hypersonic vehicle is very harsh, as the peak heat flux in this region can be even higher than that at the stagnation point. Therefore, studying the aero-heating of the rudder shaft is of great significance for designing the thermal protection system of the hypersonic vehicle. In the wind tunnel test of the aero-heating effect, we find that with the increase of the angle of attack of the lifting body model, the increasement of the heat flux of the rudder shaft is larger under laminar flow conditions than that under turbulent flow conditions. To understand this, we design a wind tunnel experiment to study the effect of laminar/turbulent hypersonic boundary layers on the heat flux of the rudder shaft under the same wind tunnel freestream conditions. The experiment is carried out in the ?2 m shock tunnel(FD-14 A) affiliated to the China Aerodynamics Research and Development Center(CARDC). The laminar boundary layer on the model is triggered to a turbulent one by using vortex generators, which are 2 mm-high diamonds. The aero-heating of the rudder shaft(with the rudder) and the protuberance(without the rudder) are studied in both hypersonic laminar and turbulent boundary layers under the same freestream condition. The nominal Mach numbers are 10 and 12, and the unit Reynolds numbers are2.4 × 10~6 m^(-1) and 2.1 × 10~6 m-1. The angle of attack of the model is 20°, and the deflection angle of the rudder and the protuberance is 10°. The heat flux on the model surface is measured by thin film heat flux sensors, and the heat flux distribution along the center line of the lifting body model suggests that forced transition is achieved in the upstream of the rudder. The test results of the rudder shaft and the protuberance show that the heat flux of the rudder shaft is lower in the turbulent flow than that in the laminar flow, but the heat flux of the protuberance is the other way around,i.e., lower in the laminar flow than in the turbulent flow. The wind tunnel test results is also validated by numerical simulations. Our analysis suggests that this phenomenon is due to the difference of boundary layer velocities caused by different thickness of boundary layer between laminar and turbulent flows, as well as the restricted flow within the rudder gap. When the turbulent boundary layer is more than three times thicker than that of the laminar boundary layer, the heat flux of the rudder shaft under the laminar flow condition is higher than that under the turbulent flow condition. Discovery of this phenomenon has great importance for guiding the design of the thermal protection system for the rudder shaft of hypersonic vehicles. | Qiang LI Liang NIE Kouli ZHANG Yu LI Suyu CHEN Guangsheng ZHU | 2019 | Chinese Journal of Aeronautics2019，32，5： | 11 |
| 18 | Deep learning applied to two-dimensional color Doppler flow imaging ultrasound images significantly improves diagnostic performance in the classification of breast masses:a multicenter study显示文摘Background:The current deep learning diagnosis of breast masses is mainly reflected by the diagnosis of benign and malignant lesions.In China,breast masses are divided into four categories according to the treatment method:inflammatory masses,adenosis,benign tumors,and malignant tumors.These categorizations are important for guiding clinical treatment.In this study,we aimed to develop a convolutional neural network(CNN)for classification of these four breast mass types using ultrasound(US)images.Methods:Taking breast biopsy or pathological examinations as the reference standard,CNNs were used to establish models for the four-way classification of 3623 breast cancer patients from 13 centers.The patients were randomly divided into training and test groups(n=1810 vs.n=1813).Separate models were created for two-dimensional(2D)images only,2D and color Doppler flow imaging(2D-CDFI),and 2D-CDFI and pulsed wave Doppler(2D-CDFI-PW)images.The performance of these three models was compared using sensitivity,specificity,area under receiver operating characteristic curve(AUC),positive(PPV)and negative predictive values(NPV),positive(LR+)and negative likelihood ratios(LR-),and the performance of the 2D model was further compared between masses of different sizes with above statistical indicators,between images from different hospitals with AUC,and with the performance of 37 radiologists.Results:The accuracies of the 2D,2D-CDFI,and 2D-CDFI-PW models on the test set were 87.9%,89.2%,and 88.7%,respectively.The AUCs for classification of benign tumors,malignant tumors,inflammatory masses,and adenosis were 0.90,0.91,0.90,and 0.89,respectively(95%confidence intervals[CIs],0.87-0.91,0.89-0.92,0.87-0.91,and 0.86-0.90).The 2D-CDFI model showed better accuracy(89.2%)on the test set than the 2D(87.9%)and 2D-CDFI-PW(88.7%)models.The 2D model showed accuracy of 81.7%on breast masses≤1 cm and 82.3%on breast masses>1 cm;there was a significant difference between the two groups(P<0.001).The accuracy of the CNN classifications for the test set(89.2%)was significantly higher than that of all the radiologists(30%).Conclusions:The CNN may have high accuracy for classification of US images of breast masses and perform significantly better than human radiologists.Trial registration:Chictr.org,ChiCTR1900021375;http://gffzz62a03ee25e6f4a77hkcufnqk60qov60wv.ffgz.tsg.suse.edu.cn/showproj.aspx?proj=33139. | Teng-Fei Yu Wen He Cong-Gui Gan Ming-Chang Zhao Qiang Zhu Wei Zhang Hui Wang Yu-Kun Luo Fang Nie Li-Jun Yuan Yong Wang Yan-Li Guo Jian-Jun Yuan Li-Tao Ruan Yi-Cheng Wang Rui-Fang Zhang Hong-Xia Zhang Bin Ning Hai-Man Song Shuai Zheng Yi Li Yang Guang | 2021 | Chinese Medical Journal2021，，4： | 11 |
| 19 | Low-temperature conversion of ammonia to nitrogen in water with ozone over composite metal oxide catalyst显示文摘As one of the most important water pollutants, ammonia nitrogen emissions have increased year by year, which has attracted people's attention. Catalytic ozonation technology, which involves production of ·OH radical with strong oxidation ability, is widely used in the treatment of organic-containing wastewater. In this work, MgO-Co_3O_4 composite metal oxide catalysts prepared with different fabrication conditions have been systematically evaluated and compared in the catalytic ozonation of ammonia(50 mg/L) in water. In terms of high catalytic activity in ammonia decomposition and high selectivity for gaseous nitrogen, the catalyst with MgO-Co_3O_4 molar ratio 8:2, calcined at 500°C for 3 hr, was the best one among the catalysts we tested, with an ammonia nitrogen removal rate of 85.2% and gaseous nitrogen selectivity of44.8%. In addition, the reaction mechanism of ozonation oxidative decomposition of ammonia nitrogen in water with the metal oxide catalysts was discussed. Moreover, the effect of coexisting anions on the degradation of ammonia was studied, finding that SO_2-4 and HCO-3 could inhibit the catalytic activity while CO_2-3 and Br-could promote it. The presence of coexisting cations had very little effect on the catalytic ozonation of ammonia nitrogen. After five successive reuses, the catalyst remained stable in the catalytic ozonation of ammonia. | Yunnen Chen Ye Wu Chen Liu Lin Guo Jinxia Nie Yu Chen Tingsheng Qiu | 2018 | Journal of Environmental Sciences2018，30，4： | 10 |
| 20 | TaqMan Real-time RT-PCR Assay for Detecting and Differentiating Japanese Encephalitis Virus显示文摘Objective To detect Japanese encephalitis virus(JEV) rapidly and distinguish its genotypes, a TaqMan-based reverse transcriptase quantitative polymerase chain reaction(RT-PCR) detection system was developed.Methods By aligning the full-length sequences of JEV(G1-G5), six sets of highly specific TaqMan real-time RT-PCR primers and probes were designed based on the highly conserved NS1, NS2, and M genes of JEV, which included one set for non-specific JEV detection and five sets for the detection of specific JEV genotypes. Twenty batches of mosquito samples were used to evaluate our quantitative PCR assay.Results With the specific assay, no other flavivirus were detected. The lower limits of detection of the system were 1 pfu/mL for JEV titers and 100 RNA copies/μL. The coefficients of variation of this real-time RT-PCR were all < 2.8%. The amplification efficiency of this method was between 90% and 103%.Conclusion A TaqMan real-time RT-PCR detection system was successfully established to detect and differentiate all five JEV genotypes. | SHAO Nan LI Fan NIE Kai FU Shi Hong ZHANG Wei Jia HE Ying LEI Wen Wen WANG Qian Ying LIANG Guo Dong CAO Yu Xi WANG Huan Yu | 2018 | Biomedical and Environmental Sciences2018，31，3： | 10 |
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