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Method for Isolating Mitochondrial DNA from Etiolated Tissue of Cabbage

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Journal of Northeast Agricultural University(English Edition) 2014年第3期21卷 23-29,F0003页

作者： Wang Shuai Wang Chao Zhang xiao-xuan College of Horticulture Northeast Agricultural University

Isolation of high-quality mitochondrial DNA（mtDNA） is an important premise for researching molecular mechanisms in cytoplasmic male sterility of cabbage（Brassica oleracea ***）. An efficient protocol for separation and purification of mitochondria and extraction of mitochondrial DNA（mtDNA） from etiolated tissues of cabbage was developed. We took a method combined mannitol density gradient with differential centrifugation, selected appropriate rotational speed, extended DNase I treating time and changed mitochondria cracking condition. The results showed that the extracted mitochondria in this protocol had complete structure, appeared to ellipsoid and had not been contaminated with other impurities under the Jannus Green B staining. The isolated mitochondrial DNA had high purity and yield through detecting the optical density, nuclear specific primer PCR and agarose gel electrophoresis. The results indicated that mitochondrial DNA extracted by this protocol had high quality and enabled to be used in futher genetic studies.

关键词： cabbage cytoplasmic male sterility(CMS) mitochondrial DNA(mtDNA) isolation

水泥搅拌桩复合地基的承载力与破坏模式研究（英文）

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Journal of Zhejiang University-Science A(Applied Physics & Engineering) 2018年第4期19卷 266-276页

作者： Hai-zuo ZHOU Gang ZHENG xiao-xuan YU Tian-qi ZHANG Jing-jin LIU School of Civil Engineering Tianjin University Key Laboratory of Coastal Civil Engineering Structure and Safety of Ministry of Education Tianjin University State Key Laboratory of Hydraulic Engineering Simulation and Safety Tianjin University

目的:工程规范中采取保守设计方法导致含有埋深的复合地基的承载力被严重低估。本文基于设计表格的方法展示水泥搅拌站复合地基的承载力系数(N_c、N_q和N_γ),分析埋深存在时的3种破坏模式,探讨承载力系数N_q和破坏模式随着各类因素(桩... 详细信息

目的:工程规范中采取保守设计方法导致含有埋深的复合地基的承载力被严重低估。本文基于设计表格的方法展示水泥搅拌站复合地基的承载力系数(N_c、N_q和N_γ),分析埋深存在时的3种破坏模式,探讨承载力系数N_q和破坏模式随着各类因素(桩长、置换率和埋置深度)变化的原因。创新点:1.确定简化的均质化水泥搅拌桩加固地基模型;2.建立非连续布局优化法(DLO)模型,计算工程实用设计表格;3.分析极限承载力系数N_q、破坏模式和各类影响因素的内在关联。方法:1.通过等效强度法确定合理的均质化数值计算模型,并与群桩模型和前人研究进行对比验证(图4和表1);2.通过极限分析上限解结合非连续布局优化法,进行大量计算,建立极限承载力系数的设计表格(图6~8)。结论:1.对于无埋深工况,承载力系数N_c随着置换率的增长而增大,直到某一临界值,此时发生实体基础破坏。2.在置换率较低时,承载力系数N_γ随着置换率的增长而减小,因为此时加固区内的等效内摩擦角减小。3.埋置深度对承载力和破坏模式产生复杂影响;当破坏模式从实体基础转化为复合型破坏时,N_q增长;随着桩长进一步增加,破坏面通过加固区内部时,N_q在减小之后保持不变。

关键词：承载力水泥搅拌桩埋置深度破坏模式

A Human t-PA Mutant cDNA Cassette Knocked in the Murine fgfr-4 Locus Targeting for Mammary Gland Expression

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Acta Biochimica et Biophysica Sinica 2004年第7期36卷 450-456页

作者： Fu-Yu LIN#, Hong-Xing CHEN#, xuan CHENG, xiao YANG*, Ji-Xian DENG*, and Pei-Tang HUANG*Beijing Institute of Biotechnology, No.20 Dongdajie Street, Fengtai District, Beijing 100071, China Beijing Institute of Biotechnology No.20 Dongdajie StreetFengtai District Beijing 100071 China

The expression of foreign gene in transgenic animals produced by pronuclear microinjection is often confounded by the position effects caused by not only the nature of chromosomal integration site but also the number and arrangement of multiple transgene copies. Gene targeting provides a new way to overcome these inhibitions by introducing single-copy transgene into a chosen site. The choice of a good chromosomal site will favor transgene expression in a predictable fashion. In this study, we tested a new site (fgfr-4) for foreign gene integration and expression. A t-PA mutant (t-PAm) expression cassette under bovine αs1-casein regulatory sequences was efficiently knocked-in fgfr-4 site through homologous recombination. The t-PAm was expressed in the milk of all targeted mice. Our experiment indicates that the fgfr-4 may be a candidate site for knocking foreign gene to make transgenic animals.

关键词： gene targeting fgfr-4 transgenic mammary gland bioreactor integration site t-PA mutant

Comparison of clinical features and outcomes in peritoneal dialysisassociated peritonitis patients with and without diabetes:A multicenter retrospective cohort study

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World Journal of Diabetes 2020年第10期11卷 435-446页

作者： Ling-Fei Meng Li-Ming Yang Xue-Yan Zhu xiao-xuan Zhang Xin-Yang Li Jing Zhao Shi-Chen Liu xiao-Hua Zhuang Ping Luo Wen-Peng Cui Department of Nephrology The Second Hospital of Jilin UniversityChangchun 130041Jilin ProvinceChina Department of Nephrology The First Hospital of Jilin University-the Eastern DivisionChangchun 130041Jilin ProvinceChina Department of Nephrology Jilin Central HospitalJilin 132011Jilin ProvinceChina Department of Nephrology Jilin FAW General HospitalChangchun 130041Jilin ProvinceChina

BACKGROUND The number of end-stage renal disease patients with diabetes mellitus(DM)who are undergoing peritoneal dialysis is *** dialysis-associated peritonitis(PDAP)is a serious complication of peritoneal dialysis leading to technical failure and increased mortality in patients undergoing peritoneal *** profile of clinical symptoms,distribution of pathogenic organisms,and response of PDAP to medical management in the subset of end-stage renal disease patients with DM have not been reported *** results have been found in long-term prognostic outcomes of PDAP in patients with *** inferred that DM is associated with bad outcomes in PDAP *** To compare the clinical features and outcomes of PDAP between patients with DM and those *** In this multicenter retrospective cohort study,we enrolled patients who had at least one episode of PDAP during the study *** patients were followed for a median of 31.1 *** were divided into a DM group and a non-DM *** features,therapeutic outcomes,and long-term prognostic outcomes were compared between the two *** factors associated with therapeutic outcomes of PDAP were analyzed using multivariable logistic regression.A Cox proportional hazards model was constructed to examine the influence of DM on patient survival and incidence of technical *** Overall,373 episodes occurred in the DM group(n=214)and 692 episodes occurred in the non-DM group(n=395).The rates of abdominal pain and fever were similar in the two groups(P>0.05).The DM group had more infections with coagulase-negative Staphylococcus and less infections with Escherichia coli(***)as compared to the non-DM group(P0.05).Patients in the DM group were older and had a higher burden o

关键词： Diabetes mellitus Mortality Peritoneal dialysis Peritoneal dialysis-associated peritonitis Technical failure

Glutamate Impairs Mitochondria Aerobic Respiration Capacity and Enhances Glycolysis in Cultured Rat Astrocytes

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Biomedical and Environmental Sciences 2017年第1期30卷 44-51页

作者： YAN Xu SHI Zhong Fang XU Li Xin LI Jia Xin WU Min WANG xiao xuan JIA Mei DONG Li Ping YANG Shao Hua YUAN Fang Department of Pathophysiology Beijing Neurosurgical Institute Beijing Tiantan Hospital Capital Medical University Beijing 100050 China Beijing Key Laboratory of Central Nervous System Injury Beijing Neurosurgical Institute Beijing Tiantan Hospital Capital Medical University Beijing 100050 China Beijing Key Laboratory of Translational Medicine for Cerebrovascular Disease Beo'ing Tiantan Hospital Capital Medical University Beijing 100050 China China National Clinical Research Center for Neurological Diseases Beijing Tiantan Hospital Capital Medical University Beijing 100050 China Department of Pharmacology and Neuroscience University of North Texas' Health Science Center Fort Worth Texas 76107-2699 USA

Objective To study the effect of glutamate on metabolism, shifts in glycolysis and lactate release in rat astrocytes. Methods After 10 days, secondary cultured astrocytes were treated with 1 mmol/L glutamate for 1 h, and the oxygen consumption rates （OCR） and extra cellular acidification rate （ECAR） was analyzed using a Seahorse XF 24 Extracellular Flux Analyzer. Cell viability was then evaluated by MTT assay. Moreover, changes in extracellular lactate concentration induced by glutamate were tested with a lactate detection kit. Results Compared with the control group, treatment with 1 mmol/L glutamate decreased the astrocytes’ maximal respiration and spare respiratory capacity but increased their glycolytic capacity and glycolytic reserve. Further analysis found that 1-h treatment with different concentrations of glutamate （0.1-1 mmol/L） increased lactate release from astrocytes, however the cell viability was not affected by the glutamate treatment. Conclusion The current study provided direct evidence that exogenous glutamate treatment impaired the mitochondrial respiration capacity of astrocytes and enhanced aerobic glycolysis, which could be involved in glutamate injury or protection mechanisms in response to neurological disorders.

关键词： Astrocytes Glutamate Mitochondrial metabolism Glycolysis Lactate

Risk factors for mortality within 6 mo in patients with diabetes undergoing urgent-start peritoneal dialysis:A multicenter retrospective cohort study

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World Journal of Diabetes 2022年第4期13卷 376-386页

作者： Si-Yu Cheng Li-Ming Yang Zhan-Shan Sun xiao-xuan Zhang Xue-Yan Zhu Ling-Fei Meng Shi-Zheng Guo xiao-Hua Zhuang Ping Luo Wen-Peng Cui Department of Nephrology The Second Hospital of Jilin UniversityChangchun 130041Jilin ProvinceChina Department of Nephrology The First Hospital of Jilin University-the Eastern DivisionChangchun 130041Jilin ProvinceChina Department of Nephrology Xing’anmeng People’s HospitalUlan Hot 137400Inner Mongolia Autonomous Region China Department of Nephrology Jilin FAW General HospitalChangchun 130041Jilin ProvinceChina Department of Nephrology Jilin Central HospitalJilin 132011Jilin ProvinceChina

BACKGROUND The risk of early mortality of patients who start dialysis urgently is high;however,in patients with diabetes undergoing urgent-start peritoneal dialysis(USPD),the risk of,and risk factors for,early mortality are *** To identify risk factors for mortality during high-risk periods in patients with diabetes undergoing *** This retrospective cohort study enrolled 568 patients with diabetes,aged≥18 years,who underwent USPD at one of five Chinese centers between 2013 and *** divided the follow-up period into two survival phases:The first 6 mo of USPD therapy and the months *** compared demographic and baseline clinical data of living and deceased patients during each ***-Meier survival curves were generated for all-cause mortality according to the New York Heart Association(NYHA)classification.A multivariate Cox proportional hazard regression model was used to identify risk factors for mortality within the first 6 mo and after 6 mo of *** Forty-one patients died within the first 6 mo,accounting for the highest proportion of mortalities(26.62%)during the entire follow-up *** disease was the leading cause of mortality within 6 mo(26.83%)and after 6 mo(31.86%).The risk of mortality not only within the first 6 mo but also after the first 6 mo was higher for patients with obvious baseline heart failure symptoms than for those with mild or no heart failure *** risk factors for mortality within the first 6 mo were advanced age hazard ratio(HR:1.908;95%CI:1.400-2.600;P<0.001),lower baseline serum creatinine level(HR:0.727;95%CI:0.614-0.860;P<0.001),higher baseline serum phosphorus level(HR:3.162;95%CI:1.848-5.409;P<0.001),and baseline NYHA class III-IV(HR:2.148;95%CI:1.063-4.340;P=0.033).Independent risk factors for mortality after 6 mo were advanced age(HR:1.246;95%CI:1.033-1.504;P=0.022)and baseline NYHA class III-IV(HR:2.015;95%CI:1.298-3.130;P=0.002).CONCLUSION To reduce the risk

关键词： Peritoneal dialysis Urgent start Diabetes mellitus,Mortality within the first 6 mo Risk factor End-stage renal disease

Detachment-Independent Cationic-Dipeptide Beacons:Reduced False-Negative Signal and Accelerated Fluorescent Lighting in Cell Imaging

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Journal of Analysis and Testing 2022年第1期6卷 19-26页

作者： Guang-Wei Fu Guo-Yan Luo xiao-xuan Li Zhen Zou Yan-Li Lei Zhi-He Qing Jue-Wen Liu Rong-Hua Yang Hunan Provincial Key Laboratory of Cytochemistry School of Chemistry and Chemical EngineeringChangsha University of Science and TechnologyChangsha 410114China College of Chemistry and Chemical Engineering Hunan Normal UniversityChangsha 410081China Department of Chemistry Waterloo Institute for NanotechnologyUniversity of WaterlooWaterlooON N2L 3G1Canada

Nanocarriers have been developed to deliver DNA probes into cells for imaging analysis and improving their nuclease ***,we found obvious lowered sensitivity and even false-negative results in the traditional DNA detachment-dependent fluorescent lighting ***,we developed a detachment-independent fluorescent lighting mechanism integrated with cationic dipeptide nanoparticles(CDNs).CDNs displayed little quenching effect on fluorophores labeled on probes and improved the nuclease resistance of *** contrast to the traditional beacon,the fluorescence lighting was significantly accelerated without the need of desorption of the recognition products from the nanosurface,avoiding false-negative results from non-specific conformational *** work will open up new thoughts to improve the sensing performance of DNA probes in cell imaging,such as speed and sensitivity.

关键词： Cationic dipeptide nanoparticles DNA probe False-negative signal Fluorescence Cell imaging

Stability of a type 2 diabetes rat model induced by high-fat diet feeding with low-dose streptozotocin injection

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2018年第7期19卷 559-569页

作者： xiao-xuan GUO Yong WANG Kai WANG Bao-ping JI Feng ZHOU Beijing Key Laboratory of Functional Food from Plant Resources College of Food Science and Nutritional Engineering China Agricultural University Beijing 100083 China Academy of State Administration of Grain Beijing 100037 China Institute of Apicultural Research Chinese Academy of Agricultural Sciences Beijing 100093 China

Objective： The present study aims at determining the stability of a popular type 2 diabetes rat model induced by a high-fat diet combined with a low-dose streptozotocin injection. Methods： Wistar rats were fed with a high-fat diet for 8 weeks followed by a one-time injection of 25 or 35 mg/kg streptozotocin to induce type 2 diabetes. Then the diabetic rats were fed with regular diet/high-fat diet for 4 weeks. Changes in biochemical parameters were monitored during the 4 weeks. Results： All the rats developed more severe dyslipidemia and hepatic dysfunction after streptozotocin injection. The features of 35 mg/kg streptozotocin rats more resembled type 1 diabetes with decreased body weight and blood insulin. Rats with 25 mg/kg streptozotocin followed by normal diet feeding showed normalized blood glucose level and pancreatic structure, indicating that normal diet might help recovery from certain symptoms of type 2 diabetes. In comparison, diabetic rats fed with high-fat diet presented decreased but relatively stable blood glucose level, and this was significantly higher than that of the control group（P〈0.05）. Conclusions： This model easily recovers with normal diet feeding. A high-fat diet is suggested as the background diet in future pharmacological studies using this model.

关键词： High-fat diet Stability Streptozotocin Type 2 diabetes mellitus

Construction of Genetic Linkage Map Based on SSR Markers in Peanut(Arachis hypogaea L.)

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Agricultural Sciences in China 2008年第8期7卷 915-921页

作者： HONG Yan-bin LIANG xuan-qiang CHEN xiao-ping LIU Hai-yan ZHOU Gui-yuan LI Shao-xiong WEN Shi-jie Crops Research Institute Guangdong Academy of Agricultural Sciences Guangzhou 510640 P.R.China

Molecular genetic maps of crop species can be used in a variety of ways in breeding and genomic research such as identification and mapping of genes and quantitative trait loci （QTLs） for morphological, physiological and economic traits of crop species. However, a comprehensive genetic linkage map for cultivated peanut has not yet been developed due to the extremely low frequency of DNA polymorphism in cultivated peanut. In this study, 142 recombinant inbred lines （RILs） derived from a cross between Yueyou 13 and Zhenzhuhei were used as mapping population in peanut （Arachis hypogaea L.）. A total 652 pairs of genomic-SSR primer and 392 pairs of EST-SSR primer were used to detect the polymorphisms between the two parents. 141 SSR primer pairs, 127 genomic-SSR and 14 EST-SSR ones, which can be used to detect polymorphisms between the two parents, were selected to analyze the RILs population. Thus, a linkage genetic map which consists of 131 SSR loci in 20 linkage groups, with a coverage of 679 cM and an average of 6.12 cM of inter-maker distance was constructed. The putative functions of 12 EST-SSR markers located on the map were analyzed. Eleven showed homology to gene sequences deposited in GenBank. This is the first report of construction of a comprehensive genetic map with SSR markers in peanut （Arachis hypogaea L.）. The map presented here will provide a genetic framework for mapping the qualitative and quantitative trait in peanut.

关键词： peanut (Arachis hypogaea L.) SSR genetic linkage map