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Identifying the stability of housekeeping genes to be used for the quantitative real-time PCR normalization in retinal tissue of streptozotocin-induced diabetic rats
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作者 Muhammad Zulfiqah Sadikan Nurul Alimah Abdul Nasir +2 位作者 Mohammad Johari Ibahim Igor Iezhitsa Renu Agarwal 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2024年第5期794-805,共12页
AIM:To investigate the stability of the seven housekeeping genes:beta-actin(ActB),glyceraldehyde-3-phosphate dehydrogenase(GAPDH),18s ribosomal unit 5(18s),cyclophilin A(CycA),hypoxanthine-guanine phosphoribosyl trans... AIM:To investigate the stability of the seven housekeeping genes:beta-actin(ActB),glyceraldehyde-3-phosphate dehydrogenase(GAPDH),18s ribosomal unit 5(18s),cyclophilin A(CycA),hypoxanthine-guanine phosphoribosyl transferase(HPRT),ribosomal protein large P0(36B4)and terminal uridylyl transferase 1(U6)in the diabetic retinal tissue of rat model.METHODS:The expression of these seven genes in rat retinal tissues was determined using real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR)in two groups;normal control rats and streptozotocininduced diabetic rats.The stability analysis of gene expression was investigated using geNorm,NormFinder,BestKeeper,and comparative delta-Ct(ΔCt)algorithms.RESULTS:The 36B4 gene was stably expressed in the retinal tissues of normal control animals;however,it was less stable in diabetic retinas.The 18s gene was expressed consistently in both normal control and diabetic rats’retinal tissue.That this gene was the best reference for data normalisation in RT-qPCR studies that used the retinal tissue of streptozotocin-induced diabetic rats.Furthermore,there was no ideal gene stably expressed for use in all experimental settings.CONCLUSION:Identifying relevant genes is a need for achieving RT-qPCR validity and reliability and must be appropriately achieved based on a specific experimental setting. 展开更多
关键词 housekeeping genes stability real-time reverse transcription polymerase chain reaction retinal tissue streptozotocin-induced diabetic rats
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Reverse water gas shift reaction over Co-precipitated Ni-CeO_2 catalysts 被引量:13
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作者 王路辉 张少星 刘源 《Journal of Rare Earths》 SCIE EI CAS CSCD 2008年第1期66-70,共5页
The Ni-CeO2 catalysts with different Ni contents were prepared by a co-precipitation method and used for Reverse Water Gas Shift (RWGS) reaction. 2wt.%Ni-CeO2 showed excellent catalytic performance in terms of activ... The Ni-CeO2 catalysts with different Ni contents were prepared by a co-precipitation method and used for Reverse Water Gas Shift (RWGS) reaction. 2wt.%Ni-CeO2 showed excellent catalytic performance in terms of activity, selectivity, and stability for RWGS reaction. Characterizations of the catalyst samples were conducted by XRD and TPR. The results indicated that, in Ni-CeO2 catalysts, there were three kinds of nickel, nickel ions in ceria lattice, highly dispersed NiO and bulk NiO. Oxygen vacancies were formed in CeO2 lattice due to the incorporation of Ni^2+ ions into ceria lattice. Oxygen vacancies formed in ceria lattice and highly dispersed Ni were key active components for RWGS, and bulk Ni was key active component for methanation of CO2. 展开更多
关键词 reverse water gas shift reaction NICKEL CEO2 oxygen vacancy
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Molybdenum carbide clusters for thermal conversion of CO2 to CO via reverse water-gas shift reaction 被引量:4
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作者 Ying Ma Zhanglong Guo +3 位作者 Qian Jiang Kuang-Hsu Wu Huimin Gong Yuefeng Liu 《Journal of Energy Chemistry》 SCIE EI CAS CSCD 2020年第11期37-43,共7页
Molybdenum carbides are highly active for CO2 conversion to CO via the reverse water-gas shift(RWGS)reaction, however the large grain size up to micrometers renders its relatively lower active sites utilization effici... Molybdenum carbides are highly active for CO2 conversion to CO via the reverse water-gas shift(RWGS)reaction, however the large grain size up to micrometers renders its relatively lower active sites utilization efficiency while generating CH4 as a by-product. In this work, a homogeneously dispersed molybdenum carbide hybrid catalyst with sub-nanosized cluster(the average size as small as 0.5 nm) is prepared via a facile carbothermal treatment for highly selective CO2-CO reduction. The partially disordered Mo2C clusters are characterized by synchrotron high-resolution XRD and atomic resolution HAADF-STEM analysis, for which the source cause of the disorder is pinpointed by XAFS analysis to be the nitrogen intercalants from the carbonaceous precursor. The partially disordered Mo2C clusters show a RWGS rate as high as 184.4 μmol gMo2C-1s-1 at 400 ℃ with a superior selectivity toward CO(> 99.5%). This work 2 highlights a facile strategy for fabricating highly dispersed and partially disordered Mo2C clusters at a sub-nano size with beneficial N-doping for delivering high catalytic activity and operational stability. 展开更多
关键词 Co2 conversion Reverse water-gas-shift reaction Molybdenum carbide cluster Sub-nanosize CO selectivity
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Detection of circulating hepatocellular carcinoma cells in peripheral venous blood by reverse transcription-polymerase chain reaction 被引量:5
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作者 Yang Liu Meng-Chao Wu +1 位作者 Guang-Xiang Qian Bai-He Zhang From the Institute of East Hepatobiliary Surgery, Second Military Medical University, Shanghai 200438, China 《Hepatobiliary & Pancreatic Diseases International》 SCIE CAS 2002年第1期72-76,共5页
Objective: To detect circulating hepatocellular carcino-ma by demonstrating hepatocellular carcinoma cells orhepatocyte-associated mRNA in the nuclear cell com-ponent of peripheral blood (PBL).Methods: Peripheral bloo... Objective: To detect circulating hepatocellular carcino-ma by demonstrating hepatocellular carcinoma cells orhepatocyte-associated mRNA in the nuclear cell com-ponent of peripheral blood (PBL).Methods: Peripheral blood (5 ml) samples were ob-tained from 93 patients with hepatocellular carcinoma(HCC) and from 33 control subjects (9 with liver cir-rhosis after hepatitis B,14 with chronic hepatitis B,10with normal liver function). To identify HCC cells inperipheral blood, liver-specific human alpha-fetopro-tein (AFP) mRNA was amplified from total RNA ex-tracted from whole blood by reverse transcription-polymerase chain reaction.Results: AFPmRNA was detected in 50 blood samplesfrom the HCC patients (50/93, 53.8%). In contrast,there were no clinical control patients whose samplesshowed detectable AFPmRNA in PBL. The presence ofAFPmRNA in blood seemed to be correlated with thestage (by TNM classification) of HCC, the serum AFPvalue, and the presence of intrahepatic metastasis,portal vein thrombosis, tumor diameter and/or distantmetastasis. In addition, AFPmRNA was detected in theblood of 21 patients with metastasis at extrahepaticorgans (100%) in contrast to 29 (40.3%)of 72 pa-tients without metastasis.Conclusion: The presence of AFPmRNA in peripheralblood may be an indicator of malignant hepatocytes,which might predict hematogenous spreading metasta-sis of tumor cells in patients with HCC. 展开更多
关键词 liver neoplasms ALPHA-FETOPROTEIN MRNA reverse transcription-polymerase chain reaction
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AN ENGINEERING MODEL FOR MULTICOMPONENT REVERSIBLE REACTION NETWORK
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作者 朱开宏 陈敏恒 袁渭康 《Chinese Journal of Chemical Engineering》 SCIE EI CAS 1985年第1期48-62,共15页
The treatment of a multicomponent reversible reaction network is extremely complicated because largenumber of rate constants must be precisely determined and because the calculation based on these rateconstants is ted... The treatment of a multicomponent reversible reaction network is extremely complicated because largenumber of rate constants must be precisely determined and because the calculation based on these rateconstants is tedious.In order to reduce the degrees of freedom of the process,the authors propose a methodin which the reactor and the separator are regarded as a whole.Based on this approach,an N-componentreversible reaction system can be dealt with as a two—component system.Consequently,a simple and ac-cessible way of the apparent rate determination is suggested.For fiist-order reactions,an explicit,simplifiedexpression has been derived for both lumped and distributed parameter reaction systems. 展开更多
关键词 CSTR ENG AN ENGINEERING MODEL FOR MULTICOMPONENT REVERSIBLE reaction NETWORK CR IND
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A PRELIMINARY STUDY ON THE THEORY OF THE SECOND AND THIRD ORDER DERIVATIVE ADSORPTION CHRONOPOTENTIOMETRY FOR A REVERSIBLE REACTION
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作者 Hong Guang ZHANG Wen Rui JIN Department of Chemistry,Shandong University,Jinan,250100,ShandongFu Song HUANG Department of Computer Science,Shandong University,Jinan,250100,Shandong 《Chinese Chemical Letters》 SCIE CAS CSCD 1993年第3期257-258,共2页
The equations of the second and third order derivative curves of time with respect to potential for a reversible process in adsorption chronopotentiometry are derived and experimentally verified.
关键词 In RT A PRELIMINARY STUDY ON THE THEORY OF THE SECOND AND THIRD ORDER DERIVATIVE ADSORPTION CHRONOPOTENTIOMETRY FOR A REVERSIBLE reaction
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COMBINED DETECTION OF BREAST CANCER MICROMETASTASES IN THE LYMPH NODES AND BONE MARROW USING REVERSETRANSCRIPTASE CHAIN REACTION AND SOUTHERN HYBRIDIZATION
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作者 李金锋 张蕾 +2 位作者 孙素莲 徐光炜 林本耀 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2000年第1期29-34,共6页
Objective: The presence of lymph nodes and bone marrow micrometastases of patients with breast carcinoma by immunohistochemistry (IHC) methods has been strongly correlated to early recurrence and shorter overall survi... Objective: The presence of lymph nodes and bone marrow micrometastases of patients with breast carcinoma by immunohistochemistry (IHC) methods has been strongly correlated to early recurrence and shorter overall survival. The aim of this study was to detect micrometastases in matched sample pairs of lymph nodes and the bone marrow of primary breast cancer patients using a more sensitive method, and compare with other clinical parameters. Methods: Cytokeratin 19 (CK-19) gene mRNA expression was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot hybridization. Human breast cancer cell line T47D was mixed with bone marrow cells at different proportions. The positive detection rate was compared among RT-PCR, Southern blotting and IHC methods. Results: Cytokeratin 19 gene was expressed in all 6 positive control samples, while the expression wasn’t seen in 18 negative control samples. CK-19 IHC positive cells were detected at a dilution of one T47D cell in 5×105 bone marrow cells, while the sensitivity detected by PCR and Southern blot hybridization was at 1:5×104 and 1:106, respectively. In the samples from the 35 patients, we found CK-19 positive cells in 2 cases (5.7%) by IHC. CK-19 gene expression signal was detected in 14/35 (40%) by RT-PCR, and 17/35 (48.6%) by southern blotting. Four cases were micrometastases positive both in lymph node and bone marrow (11.4%). There was no correlation between CK-19 detection and other clinical parameters. Conclusion: combined detection of micrometastases in lymph node and bone marrow by RT-PCR and Southern blotting, using CK-19 as a biological marker, is a highly sensitive method for breast cancer. 展开更多
关键词 MICROMETASTASES Cytokeratin 19 Breast cancer Reverse transcriptase-chain reaction Southern blot hybridization
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Quantification of Porcine Follicle-stimulating Hormone Receptor Messenger Ribonucleic Acid by Reverse Transcription competitive Polymerase Chain Reaction
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作者 朱长虹 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2000年第3期177-182,共6页
An easy and reliable method was developed for construction and quantification of competitive templates, which shared the same sequence as the amplified target DNA except for a 20 bp insertion in the middle by recombi... An easy and reliable method was developed for construction and quantification of competitive templates, which shared the same sequence as the amplified target DNA except for a 20 bp insertion in the middle by recombinant polymerase chain reaction (PCR). Among the advantages of competitive PCR is that any predictable or unpredictable variable that affects amplification has the same effect on both target and competitor species and that the final ratio of amplified products reflects exactly the initial targets. The utilization of a thermostable reverse transcriptase in the RT step was proposed to overcome the problem of the efficiency of target cDNA synthesis. In addition, to obtain reliable measurements, it was recommended to perform four PCR with amounts of competitive template flanking the concentration of the target mRNA. 展开更多
关键词 follicle stimulating hormone receptor MRNA reverse transcription competitive polymerase chain reaction
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Exogenous reference gene normalization for real-time reverse transcription-polymerase chain reaction analysis under dynamic endogenous transcription
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作者 Stephen Johnston Zachary Gallaher Krzysztof Czaja 《Neural Regeneration Research》 SCIE CAS CSCD 2012年第14期1064-1072,共9页
Quantitative real-time reverse transcription-polymerase chain reaction (qPCR) is widely used to investigate transcriptional changes following experimental manipulations to the nervous system. Despite the widespread ... Quantitative real-time reverse transcription-polymerase chain reaction (qPCR) is widely used to investigate transcriptional changes following experimental manipulations to the nervous system. Despite the widespread utilization of qPCR, the interpretation of results is marred by the lack of a suitable reference gene due to the dynamic nature of endogenous transcription. To address this inherent deficiency, we investigated the use of an exogenous spike-in mRNA, luciferase, as an internal reference gene for the 2ct normalization method. To induce dynamic transcription, we systemically administered capsaicin, a neurotoxJn selective for C-type sensory neurons expressing the TRPV-1 receptor, to adult male Sprague-Dawley rats. We later isolated nodose ganglia for qPCR analysis with the reference being either exogenous luciferase mRNA or the commonly used endogenous reference 13-111 tubulin. The exogenous luciferase mRNA reference clearly demonstrated the dynamic expression of the endogenous reference. Furthermore, variability of the endogenous reference would lead to misinterpretation of other genes of interest. In conclusion, traditional reference genes are often unstable under physiologically normal situations, and certainly unstable following the damage to the nervous system. The use of exogenous spike-in reference provides a consistent and easily implemented alternative for the analysis of qPCR data. 展开更多
关键词 exogenous reference gene sensory ganglia reverse transcription-polymerase chain reaction normalization INJURY neural regeneration
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Positive reverse transcription-polymerase chain reaction assay results in patients recovered from COVID-19: Report of two cases
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作者 Ke-Xin Huang Cheng He +4 位作者 Yan-Li Yang Di Huang Zhi-Xia Jiang Bang-Guo Li Heng Liu 《World Journal of Clinical Cases》 SCIE 2021年第12期2816-2822,共7页
BACKGROUND Coronavirus disease 2019(COVID-19)has spread around the globe.On February 28,2020,the World Health Organization adjusted the risk of spread and impact of COVID-19 to“very high”at the global level.Studies ... BACKGROUND Coronavirus disease 2019(COVID-19)has spread around the globe.On February 28,2020,the World Health Organization adjusted the risk of spread and impact of COVID-19 to“very high”at the global level.Studies have mainly focused on the etiology,epidemiology,and treatment of COVID-19 to limit further spread and the negative impact of the disease,while less attention has been devoted to the follow-up and reexamination of patients who recovered from COVID-19 or were released from quarantine.CASE SUMMARY This study reports two cases where patients who had negative reverse transcription-polymerase chain reaction(RT-PCR)test results and met the criteria for discharge subsequently had positive RT-PCR test results.The clinical manifestations and computed tomography(CT)findings of these patients were examined.The conversion of RT-PCR test results in these two patients may be related to false-negative and false-positive outcomes of the test.CT images helped track improvement of pulmonary lesions.CONCLUSION The timing of discharge of COVID-19 patients should be determined by comprehensive analysis of CT images and RT-PCR test results. 展开更多
关键词 COVID-19 False negative RECOVERY Reverse transcription-polymerase chain reaction SARS-CoV-2 Case report
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Selective synthesis of carbon monoxide via formates in reverse water–gas shift reaction over alumina-supported gold catalyst
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作者 Nobuhiro Ishito Kenji Hara +1 位作者 Kiyotaka Nakajima Atsushi Fukuoka 《Journal of Energy Chemistry》 SCIE EI CAS CSCD 2016年第2期304-308,共5页
Thermal decomposition of formic acid on SiO2, CeO2 and γ-Al2O3 was studied as an elementary step of reverse water–gas shit reaction(RWGS) over supported Au catalysts. γ-Al2O3 showed the highest CO selectivity amo... Thermal decomposition of formic acid on SiO2, CeO2 and γ-Al2O3 was studied as an elementary step of reverse water–gas shit reaction(RWGS) over supported Au catalysts. γ-Al2O3 showed the highest CO selectivity among the tested oxides in the decomposition of formic acid. Infrared spectroscopy showed the formation of four formate species on γ-Al2O3: three η~1-type and one μ~2-type species, and these formates decomposed to CO at 473 K or higher. Au-loaded γ-Al2O3 samples were prepared by a depositionprecipitation method and used as catalysts for RWGS. The supported Au catalyst gave CO with high selectivity over 99% from CO2 and H2, which is attributed to the formation of formates on Au and subsequent decomposition to CO on γ-Al2O3. 展开更多
关键词 Reverse water–gas shift reaction Carbon dioxide Hydrogen Formate Carbon monoxide Alumina Gold
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Exploring the regulatory mechanism of tRNA-derived fragments 36 in acute pancreatitis based on small RNA sequencing and experiments 被引量:1
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作者 Xi-Rui Fan Yun Huang +4 位作者 Yu Su Si-Jin Chen Yu-Lu Zhang Wei-Kang Huang Hui Wang 《World Journal of Gastroenterology》 SCIE CAS 2023年第30期4642-4656,共15页
BACKGROUND Acute pancreatitis(AP)is a disease featuring acute inflammation of the pancreas and histological destruction of acinar cells.Approximately 20%of AP patients progress to moderately severe or severe pancreati... BACKGROUND Acute pancreatitis(AP)is a disease featuring acute inflammation of the pancreas and histological destruction of acinar cells.Approximately 20%of AP patients progress to moderately severe or severe pancreatitis,with a case fatality rate of up to 30%.However,a single indicator that can serve as the gold standard for prognostic prediction has not been discovered.Therefore,gaining deeper insights into the underlying mechanism of AP progression and the evolution of the disease and exploring effective biomarkers are important for early diagnosis,progression evaluation,and precise treatment of AP.AIM To determine the regulatory mechanisms of tRNA-derived fragments(tRFs)in AP based on small RNA sequencing and experiments.METHODS Small RNA sequencing and functional enrichment analyses were performed to identify key tRFs and the potential mechanisms in AP.Reverse transcription quantitative polymerase chain reaction(RT-qPCR)was conducted to determine tRF expression.AP cell and mouse models were created to investigate the role of tRF36 in AP progression.Lipase,amylase,and cytokine levels were assayed to examine AP progression.Ferritin expression,reactive oxygen species,malondialdehyde,and ferric ion levels were assayed to evaluate cellular ferroptosis.RNA pull down assays and methylated RNA immunoprecipitation were performed to explore the molecular mechanisms.RESULTS RT-qPCR results showed that tRF36 was significantly upregulated in the serum of AP patients,compared to healthy controls.Functional enrichment analysis indicated that target genes of tRF36 were involved in ferroptosisrelated pathways,including the Hippo signaling pathway and ion transport.Moreover,the occurrence of pancreatic cell ferroptosis was detected in AP cells and mouse models.The results of interference experiments and AP cell models suggested that tRF-36 could promote AP progression through the regulation of ferroptosis.Furthermore,ferroptosis gene microarray,database prediction,and immunoprecipitation suggested that tRF-36 accelerated the progression of AP by recruiting insulin-like growth factor 2 mRNA binding protein 3(IGF2BP3)to the p53 mRNA m6A modification site by binding to IGF2BP3,which enhanced p53 mRNA stability and promoted the ferroptosis of pancreatic follicle cells.CONCLUSION In conclusion,regulation of nuclear pre-mRNA domain-containing protein 1B promoted AP development by regulating the ferroptosis of pancreatic cells,thereby acting as a prospective therapeutic target for AP.In addition,this study provided a basis for understanding the regulatory mechanisms of tRFs in AP. 展开更多
关键词 Acute pancreatitis tRNA-derived fragments tRNA-derived fragments 36 Mouse models Ferroptosis Reverse transcription quantitative polymerase chain reaction
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Correlation of serum SARS-CoV-2 IgM and IgG serology and clinical outcomes in COVID-19 patients:Experience from a tertiary care centre
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作者 Mohan Suresh Pratap Kumar +5 位作者 Prasan Kumar Panda Vikram Jain Rohit Raina Sarama Saha Subbiah Vivekanandhan Balram Ji Omar 《World Journal of Biological Chemistry》 2023年第2期52-61,共10页
BACKGROUND The severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)virus has become a pandemic for the last 2 years.Inflammatory response to the virus leads to organ dysfunction and death.Predicting the severit... BACKGROUND The severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)virus has become a pandemic for the last 2 years.Inflammatory response to the virus leads to organ dysfunction and death.Predicting the severity of inflammatory response helps in managing critical patients using serology tests IgG and IgM.AIM To investigate the correlation of the serology(IgM and IgG)with reverse transcriptase polymerase chain reaction(RT-PCR)status,disease severity[mild to critical],intensive care unit(ICU)admission,septic shock,acute kidney injury,and in-hospital mortality.METHODS We conducted a longitudinal study to correlate serum SARS-CoV-2 immunoglobulin M(IgM)and immunoglobulin G(IgG)serology with clinical outcomes in coronavirus disease 2019(COVID-19)patients.We analyzed patient data from March to December 2020 for those who were admitted at All India Institute of Medical Sciences Rishikesh.Clinical and laboratory data of these patients were collected from the e-hospital portal and analyzed.A correlation was seen with clinical outcomes and was assessed using MS Excel 2010 and SPSS software.RESULTS Out of 494 patients,the mean age of patients was 48.95±16.40 years and there were more male patients in the study(66.0%).The patients were classified as mild-moderate 328(67.1%),severe 131(26.8%),and critical 30(6.1%).The mean duration from symptom onset to serology testing was 19.87±30.53 d.In-hospital mortality was observed in 25.1%of patients.The seropositivity rate(i.e.,either IgG or IgM>10 AU)was 50%.IgM levels(AU/mL)(W=33428.000,P≤0.001)and IgG levels(AU/mL)(W=39256.500,P≤0.001),with the median IgM/IgG levels(AU/mL),were highest in the RT-PCR-Positive group compared to RT-PCR-Negative clinical COVID-19.There was no significant difference between the two groups in terms of all other clinical outcomes(disease severity,septic shock,ICU admission,mechanical ventilation,and mortality).CONCLUSION The study showed that serology levels are high in RT-PCR positive group compared to clinical COVID-19.However,serology cannot be useful for the prediction of disease outcomes.The study also highlights the importance of doing serology at a particular time as antibody titers vary with the duration of the disease.In week intervals there was a significant correlation between clinical outcomes and serology on week 3. 展开更多
关键词 Inflammatory response Reverse transcription polymerase chain reaction SARS-CoV-2 Serology IgM and IgG
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Structural and Electrochemical Performance of Additives-doped a-Ni(OH)_2
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作者 张仲举 朱燕娟 《Journal of Wuhan University of Technology(Materials Science)》 SCIE EI CAS 2012年第3期538-541,共4页
The additives-doped α-nickel hydroxides were prepared by supersonic co-precipitation method. The crystal structure and grain size of the prepared samples were characterized by X-ray diffraction (XRD) and Particle s... The additives-doped α-nickel hydroxides were prepared by supersonic co-precipitation method. The crystal structure and grain size of the prepared samples were characterized by X-ray diffraction (XRD) and Particle size distribution (PSD), respectively. Cyclic voltammetry (CV) tests show that Al-Co-Y doped Ni(OH)2 has better reaction reversibility, higher proton diffusion coefficient than those of Al-Co doped Ni(OH)2. Al-Co-Y doped Ni(OH)2 also has lower charge-transfer resistance as shown by electrochemical impedance spectroscopy (EIS). Charge/discharge tests show that the discharge capacity of Al-Co-Y doped Ni(OH)2 reaches 328 mAh/g at 0.2 C and 306 mAh/g at 0.5 C, while Al-Co doped Ni(OH)2 can only discharge a capacity of 308 mAh/g at 0.2 C and 267 mAh/g at 0.5 C. 展开更多
关键词 additives-doped particle size distribution reaction reversibility proton diffusion coefficient charge-transfer resistance
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Multiplex RT-PCR-based detections of CEA, CK20 and EGFR in colorectal cancer patients 被引量:19
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作者 Aikaterini Tsouma Chrysanthi Aggeli +7 位作者 Panagiotis Lembessis George N Zografos Dimitris P Korkolis Dimitrios Pectasides Maria Skondra Nikolaos Pissimissis Anastasia Tzonou Michael Koutsilieris 《World Journal of Gastroenterology》 SCIE CAS CSCD 2010年第47期5965-5974,共10页
AIM: To develop a multiplex reverse transcription polymerase chain reaction (RT-PCR) method detecting cir-culating tumor cells in the peripheral blood of colorectal cancer (CRC) patients. METHODS: Peripheral blood sam... AIM: To develop a multiplex reverse transcription polymerase chain reaction (RT-PCR) method detecting cir-culating tumor cells in the peripheral blood of colorectal cancer (CRC) patients. METHODS: Peripheral blood samples were collected from 88 CRC patients and 40 healthy individuals from the blood donors' clinic and subsequently analyzed by multiplex RT-RCR for the expression of carcinoembryonic antigen (CEA), cytokeratin 20 (CK20) and epidermal growth factor receptor (EGFR) mRNA. The analysis involved determining the detection rates of CEA, CK20 and EGFR transcripts vs disease stage and overall survival. Median follow-up period was 19 mo (range 8-28 mo). RESULTS: Rates of CEA, CK20 and EGFR detection in CRC patients were 95.5%, 78.4% and 19.3%, respectively. CEA transcripts were detected in 3 healthy volunteer samples (7.5%), whereas all control samples were tested negative for CK20 and EGFR transcripts. The increasing number of positive detections for CEA, CK20 and EGFR transcripts in each blood sample was positively correlated with Astler-Coller disease stage (P< 0.001) and preoperative serum levels of CEA (P=0.029) in CRC patients. Data analysis using Kaplan-Meier estimator documented signif icant differences in the overall survival of the different CRC patient groups as formed according to the increasing number of positivity for CEA, CK20 and EGFR transcripts. CONCLUSION: These data suggest that multiplex RTPCR assay can provide useful information concerning disease stage and overall survival of CRC patients. 展开更多
关键词 Peripheral blood Carcinoembryonic antigen Cytokeratin 20 Epidermal growth factor receptor Multiplex reverse transcription polymerase chain reaction
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Diagnosis of nervous necrosis virus in orange-spotted grouper,Epinephelus coioides, by a rapid and convenient RT-PCR method 被引量:6
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作者 MU Yinnan LIN Kebing +1 位作者 CHEN Xinhua AO Jingqun 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2013年第10期88-92,共5页
Viral nervous necrosis (VNN) causes high mortality in marine fish, especially in the grouper, worldwide and in China. Since there is no effective vaccine or drug to deal with VNN, early detection and prevention is i... Viral nervous necrosis (VNN) causes high mortality in marine fish, especially in the grouper, worldwide and in China. Since there is no effective vaccine or drug to deal with VNN, early detection and prevention is important to block its outbreak. In this study, a reverse transcription-polymerase chain reaction (RT-PCR) was developed for the rapid, convenient, and sensitive detection of the VNN pathogen, nervous necro- sis virus (NNV), in the grouper. The whole process was completed within 3.5 h from the RNA extraction to PCR product visualization. The detection limit of this method was 200 copies of NNV RNA standard, which corresponded to 200 copies of virus particles. This RT-PCR method was specific to the NNV detection with no cross-reactivity to other fish viral disease pathogens, such as infectious pancreatic necrosis virus (IPNV), infectious hematopoietic necrosis virus (IHNV), spring viraemia of carp virus (SVCV), epizootic haematopoietic necrosis virus (EHNV), and large yellow croaker iridovirus (LYC1V). With this method, the orange-spotted grouper (Epinephelus coioides) fry from hatcheries with or without incidence of the VN- N epidemic in Fujian Province were detected. The results showed that all or 93% of the fry from the two hatcheries with incidence of the epidemic were diagnosed as positive, while 40% or 25% of fry from the t- wo hatcheries without the VNN epidemic were also detected as NNV positive, indicating that this RT-PCR method can be used for rapid, sensitive detection of NNV infection and applied in the VNN epidemic alert. 展开更多
关键词 viral nervous necrosis nervous necrosis virus orange-spotted grouper (Epinephelus coioides) reverse transcription-polymerase chain reaction DETECTION
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Diagnostic value of cancer-testis antigen mRNA in peripheral blood from hepatocellular carcinoma patients 被引量:22
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作者 Li Zhao Dong-Cheng Mou +3 位作者 Zeng-An Wu Ji-Run Peng Lei Huang Xi-Sheng Leng 《World Journal of Gastroenterology》 SCIE CAS CSCD 2010年第32期4072-4078,共7页
AIM:To evaluate the diagnostic value of cancer-testis antigen(CTA) mRNA in peripheral blood samples from hepatocellular carcinoma(HCC) patients.METHODS:Peripheral blood samples were taken from 90 patients with HCC bef... AIM:To evaluate the diagnostic value of cancer-testis antigen(CTA) mRNA in peripheral blood samples from hepatocellular carcinoma(HCC) patients.METHODS:Peripheral blood samples were taken from 90 patients with HCC before operation.Expression of melanoma antigen-1(MAGE-1),synovial sarcoma X breakpoint-1(SSX-1),and cancer-testis-associated protein of 11 kDa(CTp11) mRNA in peripheral blood mononuclear cells(PBMC) was tested by nested reverse transcriptspolymerase chain reaction(RT-PCR).Serum α-fetoprotein(AFP) in these patients was also determined.RESULTS:The positive rate of MAGE-1,SSX-1 and CTp11 transcripts was 37.7%,34.4%,31.1% in PBMC samples,and 74.4%,73.3%,62.2% in their resected tumor samples,respectively.The positive rate for at least one of the transcripts of three CTA genes was 66.7% in PBMC samples and 91.1% in their resected tumor samples.MAGE-1,SSX-1 and/or CTp11 mRNA were not detected in the PBMC of those patients from whom the resected tumor samples were MAGE-1,SSX-1 and/or CTp11 mRNA negative,nor in the PBMC samples from 20 healthy donors and 10 cirrhotic patients.Among the 90 patients,the serum AFP in 44 patients met the general diagnostic standard(AFP > 400 μg/L) for HCC,and was negative(AFP ≤ 20 μg/L) or positive with a low concentration(20 μg/L < AFP ≤ 400 μg/L) in the other patients.The positive rate for at least one of the transcripts of three CTA genes in PBMC samples from the AFP negative or positive patients with a low concentration was 69.2% and 45.0%,respectively.Of the 90 patients,71(78.9%) were diagnosed as HCC by nested RT-PCR and serum AFP.Although the positive rate for at least one of the transcripts of three CTA genes in PBMC samples from 53 patients at TNM stage or was obviously higher than that in PBMC samples from 37 patients at stage or(77.9% vs 51.4%,P = 0.010),the CTA mRNA was detected in 41.7% and 56.0% of PBMC samples from HCC patients at stages andrespectively.CONCLUSION:Detecting MAGE-1,SSX-1 and CTp11 mRNA in PBMC improves the total diagnostic rate of HCC. 展开更多
关键词 Hepatocellular carcinoma α-fetoprotein Cancer-testis antigen Diagnosis Nested reverse transcripts-polymerase chain reaction
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Identification of Sheep Endogenous Beta-Retroviruses with Uterus-Specific Expression in the Pregnant Mongolian Ewe 被引量:6
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作者 QI Jing-wei XU Meng-jie +4 位作者 LIU Shu-ying ZHANG Yu-fei LIU Yue ZHANG Ya-kun CAO Gui-fang 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2013年第5期884-891,共8页
The sheep genome harbours approximately 20 copies of endogenous beta-retroviruses (enJSRVs), and circumstantial evidence suggests that enJSRVs might play a role in mammalian reproduction, particularly placental morp... The sheep genome harbours approximately 20 copies of endogenous beta-retroviruses (enJSRVs), and circumstantial evidence suggests that enJSRVs might play a role in mammalian reproduction, particularly placental morphogenesis. This study was aimed to assess the expression of mRNAs of an enJSRV and its receptor, HYAL2, in the uterus and conceptuses of Mongolian ewes throughout gestation, using real-time reverse transcription polymerase chain reaction and in situ hybridization analysis. The results showed that enJSRV and HYAL2 mRNAs were found to be expressed throughout gestation in the endometrium, chorion, placenta, and conceptus. The enJSRV mRNA was most abundant in the placenta on day 90 of pregnancy, in the endometrium on day 30 and 50, and in the chorion on day 70 and 110. However, HYAL2 mRNA was most abundant in the endometrium on day 30. These differences were all significantly different from each other (P〈0.01). In situ hybridization showed that enJSRV and HYAL2 mRNAs were specifically expressed in endometrial luminal epithelium and glandular epithelium, trophoblastic giant binucleated cells (BNCs), endometrial caruncles, placental cotyledons, stroma, trophectoderm, as well as multinucleated syncytia of the placenta and blood vessel endothelial cells. Collectively, little is known about the molecular mechanisms by which trophoblastic differentiation and multinucleated syncytia formation are regulated by enJSRVs. However, the temporal and spatial distributions of enJSRV expression in the uterus and conceptus indicate that differentiation of BNCs and the formation of a multinucleated syncytiotrophoblast involve enJSRV and possibly its cellular receptor, HYAL2. Therefore, enJSRV and HYAL2 appear to play important roles in the female reproductive physiology in this breed of sheep. 展开更多
关键词 ENJSRV HYAL2 expression real-time reverse transcription polymerase chain reaction in situ hybridizationhybridization Mongolian ewe
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High Initial Reversible Capacity and Long Life of Ternary SnO_(2)-Co-carbon Nanocomposite Anodes for Lithium-Ion Batteries 被引量:3
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作者 Pan Deng Jing Yang +7 位作者 Shengyang Li Tian-E Fan Hong-Hui Wu Yun Mou Hui Huang Qiaobao Zhang Dong-Liang Peng Baihua Qu 《Nano-Micro Letters》 SCIE EI CAS CSCD 2019年第1期326-338,共13页
The two major limitations in the application of SnO_2 for lithium?ion battery(LIB) anodes are the large volume variations of SnO_2 during repeated lithiation/delithiation processes and a large irreversible capacity lo... The two major limitations in the application of SnO_2 for lithium?ion battery(LIB) anodes are the large volume variations of SnO_2 during repeated lithiation/delithiation processes and a large irreversible capacity loss during the first cycle, which can lead to a rapid capacity fade and unsatisfactory initial Coulombic e ciency(ICE). To overcome these limitations, we developed composites of ultrafine SnO_2 nanoparticles and in situ formed Co(CoSn) nanocrystals embedded in an N?doped carbon matrix using a Co?based metal–organic framework(ZIF?67). The formed Co additives and structural advantages of the carbon?confined SnO_2/Co nanocomposite e ectively inhibited Sn coarsening in the lithiated SnO_2 and mitigated its structural degradation while facilitating fast electronic transport and facile ionic di usion. As a result, the electrodes demonstrated high ICE (82.2%), outstanding rate capability(~ 800 mAh g^(-1) at a high current density of 5 A g^(-1)), and long?term cycling stability(~ 760 mAh g^(-1) after 400 cycles at a current density of 0.5 A g^(-1)). This study will be helpful in developing high?performance Si(Sn)?based oxide, Sn/Sb?based sulfide, or selenide electrodes for LIBs. In addition, some metal organic frameworks similar to ZIF?67 can also be used as composite templates. 展开更多
关键词 Ultrafine SnO_(2) nanostructures ZIF-67 frameworks Enhanced initial Coulombic efficiency Reversible conversion reaction
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Correlation of glycosyltransferases mRNA expression in extrahepatic bile duct carcinoma with clinical pathological characteristics 被引量:6
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作者 Xiao-Ling Jin, Shu-Sen Zheng, Bing-Sheng Wang and Hui-Li Chen Nanjing, China Department of General Surgery, Nanjing First Hospi- tal Affiliated to Nanjing Medical University, Nanjing 210006, China Department of Hepatobiliary Surgery, First Affiliated Hospital, Zhe- jiang University School of Medicine, Hangzhou 310003, China +1 位作者 Department of General Surgery, Zhongshan Hospital, Fudan Universi- ty, Shanghai 200032 , China and Key Laboratory of Glycocon- jugate Research, Ministry of Health, Department of Biochemistry, Fudan University School of Medicine, Shanghai 200032, China 《Hepatobiliary & Pancreatic Diseases International》 SCIE CAS 2004年第2期292-295,共4页
BACKGROUND: The incidence of extrahepatic bile duct carcinoma (EBDC) has been increasing, especially in aged people, but the glycobiology of the tumor is not elucida- ted. In this study we investigated the expressions... BACKGROUND: The incidence of extrahepatic bile duct carcinoma (EBDC) has been increasing, especially in aged people, but the glycobiology of the tumor is not elucida- ted. In this study we investigated the expressions of three glycosyltransferases in 35 patients with EBDC and 35 pa- tients with benign biliary duct disease (BBDD) as well as their clinicopathological significance. METHOD: The patients were divided into several sub- groups by tumor differentiation, TNM stage, and invasion by the standards recommended by UICC. Tumor samples were immediately frozen in liquid nitrogen after resection, followed by mRNA determination of enzymes in the tissue using a mRNA selective reverse trancriptase-polymerase chain reaction kit. The mRNA levels of different groups were semi-quantitatively compared. RESULTS: The mRNA levels of N-acetylglucosaminyltrans- ferase V (GnT-V) and a subtype of α2,3 sialyltransferases for N-glycans, ST3Gal- were elevated 7.75 and 5.39 times in EBDC as compared with BBDD, respectively, and they were correlated to several clinicopathological factors including tumor advancement, differentiation, metastasis, and invasiveness. The mRNA expression of another sialyl- transferase, ST6Gal- , was also 0.63-fold higher in EBDC than in BBDD, but not involved in the clinicopathological characteristics. CONCLUSION: The elevated expression of these three gly- cosyltransferases can be considered as an important molecu- lar event in the occurrence and progression of EBDC. 展开更多
关键词 extrahepatic bile duct carcinoma SIALYLTRANSFERASE N-ACETYLGLUCOSAMINYLTRANSFERASE reverse trancriptase-polymerase chain reaction
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