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The development of a rapid high-quality universal nucleic acid extraction kit based on magnetic separation

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Science China Chemistry 2017年第12期60卷 1602-1608页

作者： Bo Li Xianbo Mou Zhu Chen Hui Chen Yan Deng Song Li enben su Lei He Nongyue He State Key Laboratory of Bioelectronics Southeast UniversityNanjing 210096China The Medical School Ningbo University Ningbo 315211China Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province Hunan Key Laboratory of Green Chemistry and Application of Biological NanotechnologyHunan University of TechnologyZhuzhou 412007China Getein Biotechnology Co. Ltd.Nanjing 211505China

Nucleic acid extraction is one of the bases in molecular biology and it is the indispensable process for nucleic acid detection. Due to its excellent characteristics, the method based on magnetic beads has become one of the main methods for nucleic acid *** this research, we have developed a rapid high-quality universal nucleic acid extraction kit based on magnetic beads(MBs) and named it MB-100. Some factors affecting nucleic acid extraction were optimized, such as particle size of MBs, surface groups of MBs and lysis time. Results demonstrated that micron-grade MBs@SiO_2 particles were much more appropriate for nucleic acid extraction and lysis time was 5 min. Simultaneously, the performance of MB-100 was explored. Results showed that it had much higher extraction efficiency than other two commercial kits, and the following PCR reaction also had much higher amplification efficiency.

关键词： extraction kit MBs@SiO2 lysis time extraction efficiency

A microfluidic device for accurate detection of hs-cTnI

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Chinese Chemical Letters 2021年第4期32卷 1555-1558页

作者： Li Huang enben su Yuan Liu Nongyue He Yan Deng Lian Jin Zhu Chen Song Li State Key Laboratory of Bioelectronics National Demonstration Center for Experimental Biomedical Engineering EducationSchool of Biological Science and Medical EngineeringSoutheast UniversityNanjing 210096China Getein Biotechnology Co. Ltd.Nanjing 210000China Hunan Key Laboratory of Biomedical Nanomaterials and Devices Hunan University of TechnologyZhuzhou 412007China

This device is aimed at ensuring that the sample is uniformly and equivalently reacted with the antibody on the NC membrane in each test when the microfluidic liquid system is introduced to the *** this study,the developed microfluidic chip can avoid the presence of the sample and conjugate pads in the chip,while the precision of the chro matography system can be greatly improved using the same particles,NC membrane and antibody alongside the traditional *** results,taking the detection of cTnI as an example,revealed that the coefficient of variation(CV)is controlled within 4%,while the maximum record of the contrast chromatographic reagent strip can reach 15%.Additionally,the detection sensitivity can maintain the same order of magnitudes with that of the traditional chromatographic *** the results,the determination correlation of the developed microfluidic chip has been greatly *** addition,the CV of the chip in this study is greatly improved in comparison with that of the traditional *** biggest improvement lies in the mixing between the sample and the microspheres,indicating that this is a new approach to improve the CV of the traditional strip.

关键词： Point of care testing hs-cTnI Microfluidic device Precision

Recent biomedical applications of bio-sourced materials

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Bio-Design and Manufacturing 2018年第1期1卷 26-44页

作者： Abdelrahman Elbaz Zhenzhu He Bingbing Gao Junjie Chi enben su Dagan Zhang Songqin Liu Hua Xu Hong Liu Zhongze Gu State Key Laboratory of Bioelectronics School of Biological Science and Medical Engineering Southeast University Nanjing 210096 China Laboratory of Environment and Biosafety Research Institute of Southeast University in Suzhou S uzhou 215123 China Getein Biotech Inc. No. 9 Bofu Road Luhe District Nanjing 211505 Jiangsu China Jiangsu Engineering Laboratory of Smart Carbon-Rich Materials and Device Jiangsu Province Hi-Tech Key Laboratory for Bio-Medical Research School of Chemistry and Chemical Engineering Southeast University Nanjing 211189 China

Natural anisotropic nanostructures occurring in several organisms have gained more and more attention because of their obvious advantages in sensitivity, stability, security, miniaturization, portability, online use, and remote monitoring. Due to the development of research on nature-inspired bionic structures and the demand for highly efficient, low-cost microfabrication techniques, an understanding of and the ability to replicate the mechanism of structural coloration have become increasingly significant. These sophisticated structures have many unique functions and are used in many applications. Many sensors have been proposed based on their novel structures and unique optical properties. Several of these bio-inspired sensors have been used for infrared radiation/thermal, pH, and vapor techniques, among others, and have been discussed in detail, with an intense focus on several biomedical applications. However, many applications have yet to be discovered. In this review, we will describe these nanostructured materials based on their sources in nature and various structures, such as layered, hierarchical, and helical structures. In addition, we discuss the functions endowed by these structures, such as superhydrophobicity, adhesion, and high strength, enabling them to be employed in a number of applications in biomedical fields, including cell cultivation, biosensors, and tissue engineering.

关键词： Nature Bio-sourced Bio-inspired Bio-Materials Biomedical

Washing-free chemiluminescence immunoassay for rapid detection of cardiac troponin Ⅰ in whole blood samples

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Chinese Chemical Letters 2022年第2期33卷 743-746页

作者： Huan Zhao enben su Li Huang Yunfeng Zai Yuan Liu Zhu Chen Song Li Lian Jin Yan Deng Nongyue He State Key Laboratory of Bioelectronics National Demonstration Center for Experimental Biomedical Engineering EducationSchool of Biological Science and Medical EngineeringSoutheast UniversityNanjing 210096China Getein Biotechnology Co. Ltd.Nanjing 210000China Hunan Key Laboratory of Biomedical Nanomaterials and Devices Hunan University of TechnologyZhuzhou 412007China

Chemiluminescence immunoassay(CLⅠA) has always been a great challenge in detecting cardiac troponin Ⅰ(c Tn Ⅰ) in whole blood samples without centrifugation because of the interference of red blood cells and low sensitivity. Ⅰn this study, the antigens and erythrocytes in the blood were captured by the antibodies immobilized on the magnetic particles, recognized by another biotinconjugated c Tn Ⅰ antibody and detected by streptavidin/acridine aster-conjugated polychloromethylstyrene microspheres(PCMS). After magnetic separation, the supernatant was transferred and measured. No significant difference was noted between the c Tn Ⅰ concentrations of the serum samples,plasma samples and whole blood. The prepared PCMS provided more functional areas to conjugate streptavidin and acridinium ester, so the immunoassay has highly sensitive, the limits of blank at0.012 ng/mL, and functional sensitivity at 0.019 ng/mL with a CV of 20%, and 0.058 ng/mL with a CV of 10%. Total precision of any sample type ranged from 2.62%~5.67%. The assay was linear over the studied range of 0.01-50.00 ng/mL, and no hook effect was found when c Tn Ⅰ concentrations reached 1900 ng/mL. No significant interference was noted with the potential endogenous interfering substances. Compared with the commercial kit(Abbott assay kit), the correlation coefficient was 0.9859. A washing-free CLⅠA was established for the rapid detection of c Tn Ⅰ in human whole blood, using erythrocyte capture antibodies-conjugated magnetic nanoparticles for eliminating the influence of erythrocytes and PCMS for signal amplification, which showed great potential in clinical application.

关键词： Washing-free Whole blood Chemiluminescence immunoassay Polychloromethylstyrene microspheres

Discrimination of mitochondrial DNA 10400 locus by SNP-operated on/off Switch

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Journal of Nanjing Medical University 2008年第6期22卷 346-350页

作者： Mei Hong enben su Ziqing Chen Xiaobing Ju Qi Chen Rong Zhou The expert evidence institute the First Affiliated Hospital of Nanjing Medical University Nanjing 210029Jiangsu Province China

Objective：To apply reformed AS-PCR, which combined phosphorothioate-modified primers with exo^＋ polymerase, in single nucleotide polymorphism discrimination of mitochondrial DNA 10400 locus. Methods： We used the mtDNA 10400 locus to design unrnodifled and 3 ＇ phosphorothioate-modified allele-specific primers for PCR, which was performed using polymerases with and without 3 ＇ exonuclease activities. The effects of these primers on primer-extension were evaluated by agarose gel electrophoresis. Results： The unmodified primers were extended by both exo and exo^＋ polymerase irrespective of whether the primers were matched or mismatched with the templates. However, the 3＇ phosphorothioate-modified primers with a terminal mismatch triggered an ＂off-switch＂ of exo~ polymerase when compared to exopolymerase. Conclusion： The＂ on/off＇switch constituted by the combination of 3 ＇ phosphorothioatemodified primers with exo^＋ polymerase is a cost-effective, high-throughput and reliable method for SNP typing, which will be of enormous application in association studies by single nucleotide polymorphism screening.

关键词： SNP exo^+ polymerase phosphorothioate-modification allele-specific PCR mitochondrial DNA

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母体血浆中游离胎儿DNA研究进展

国际生殖健康／计划生育杂志 2008年第6期27卷 361-364页

作者：高慧双苏恩本南京医科大学第一附属医院检验中心 210029 南京医科大学第一附属医院司法鉴定所

母血浆中游离胎儿DNA是无创伤性产前诊断中胎儿信息的主要来源之一。近年对游离胎儿DNA的来源、生物学特点以及临床应用的研究倍受关注。现已开展胎儿性别鉴定、RhD基因检测、STR遗传标记检测、胎儿非整倍体检测等应用研究。母血浆中游... 详细信息

母血浆中游离胎儿DNA是无创伤性产前诊断中胎儿信息的主要来源之一。近年对游离胎儿DNA的来源、生物学特点以及临床应用的研究倍受关注。现已开展胎儿性别鉴定、RhD基因检测、STR遗传标记检测、胎儿非整倍体检测等应用研究。母血浆中游离胎儿DNA提取方法的发展和高灵敏度检测技术的应用,为母血浆中游离胎儿DNA的研究开辟了新前景。就母血中胎儿DNA来源、生物学特点,分析方法、基础性研究及临床应用的进展情况综述。

关键词：游离胎儿DNA 无创伤产前诊断血浆DNA 研究进展

利用微卫星多态性检测造血干细胞移植后嵌合体的研究

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青海医学院学报 2007年第4期28卷 234-239页

作者：熊辉霞冀林华苏恩本青海大学附属医院江苏省人民医院

目的利用微卫星DNA的多态性,检测造血干细胞移植后嵌合体。方法用无关个体的单个核细胞混合标本在体外模拟混合嵌合体,采用聚合酶链反应(PCR)结合聚丙烯酰胺凝胶电泳、银染色和凝胶成像分析技术半定量检测嵌合体。结果本方法的敏感性为1... 详细信息

目的利用微卫星DNA的多态性,检测造血干细胞移植后嵌合体。方法用无关个体的单个核细胞混合标本在体外模拟混合嵌合体,采用聚合酶链反应(PCR)结合聚丙烯酰胺凝胶电泳、银染色和凝胶成像分析技术半定量检测嵌合体。结果本方法的敏感性为1.25%～5%,不同位点敏感性不同;混合模拟标本扩增产物分析计算所得嵌合率与扩增前供体细胞比例呈显著直线相关(r=0.97～0.98,P<0.001),不同位点间相关性有所差异,以各位点的平均值进行线性分析更趋于直线相关(r=0.998)。结论STR-PCR结合凝胶电泳成像分析技术是一种敏感、准确、可靠、经济的嵌合体半定量检测方法。

关键词：造血嵌合体短串联重复序列造血干细胞移植

运动对大鼠肌细胞丝裂素活化蛋白酶信号传导系统的调节作用

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中华物理医学与康复杂志 2002年第6期24卷 353-355页

作者：江钟立张勤励建安周士枋陈家伟苏恩本丁小健南京医科大学第一临床医学院康复医学科 210029 南京医科大学第一临床医学院内分泌科 210029 南京医科大学第一临床医学院中心实验室 210029

目的　观察运动对心肌和骨骼肌细胞应激激活蛋白激酶 (JNK)信号通道活性的影响 ,为运动处方的制定提供分子生物学依据。方法　 1 2周龄、体重 2 0 0～ 2 30克的SD大鼠 1 8只 ,随机分为对照组 (n =6)、急性运动组 (n=6)和运动训练组 (n ... 详细信息

目的　观察运动对心肌和骨骼肌细胞应激激活蛋白激酶 (JNK)信号通道活性的影响 ,为运动处方的制定提供分子生物学依据。方法　 1 2周龄、体重 2 0 0～ 2 30克的SD大鼠 1 8只 ,随机分为对照组 (n =6)、急性运动组 (n=6)和运动训练组 (n =6)。采用活动平板 ,坡度 1 0 % ,速度 2 0m/min,运动时间 2 0分 /次。急性运动组仅运动 2 0min ;运动训练组 ,每日 1次 ,每周 5次 ,运动三周 ;对照组不参与运动。运用化学发光测定心肌和骨骼肌JNK活性。结果　骨骼肌JNK活性在急性运动组和运动训练组分别是非运动组的 4 .1倍和1 1 .1倍 ,且运动训练组较急性运动组增高 2 .71倍。心肌JNK活性在急性运动组和运动训练组分别是非运动组的 3 .5倍和 2 .1 6倍 ,且急性运动组较运动训练组增高 1 .62倍。结论　运动可以激活心肌和骨骼肌JNK信号转导通道。

关键词：运动大鼠肌细胞丝裂素活化蛋白酶信号传导系统调节作用骨骼肌