KH Profile-01
Analysis of Genetic Diversity and Population Structure
Agricultural Sciences in China2010, 9(9): 1251-1262September 2010Received 30 October, 2009 Accepted 16 April, 2010Analysis of Genetic Diversity and Population Structure of Maize Landraces from the South Maize Region of ChinaLIU Zhi-zhai 1, 2, GUO Rong-hua 2, 3, ZHAO Jiu-ran 4, CAI Yi-lin 1, W ANG Feng-ge 4, CAO Mo-ju 3, W ANG Rong-huan 2, 4, SHI Yun-su 2, SONG Yan-chun 2, WANG Tian-yu 2 and LI Y u 21Maize Research Institute, Southwest University, Chongqing 400716, P.R.China2Institue of Crop Sciences/National Key Facility for Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences,Beijing 100081, P.R.China3Maize Research Institute, Sichuan Agricultural University, Ya’an 625014, P.R.China4Maize Research Center, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100089, P.R.ChinaAbstractUnderstanding genetic diversity and population structure of landraces is important in utilization of these germplasm in breeding programs. In the present study, a total of 143 core maize landraces from the South Maize Region (SR) of China,which can represent the general profile of the genetic diversity in the landraces germplasm of SR, were genotyped by 54DNA microsatellite markers. Totally, 517 alleles (ranging from 4 to 22) were detected among these landraces, with an average of 9.57 alleles per locus. The total gene diversity of these core landraces was 0.61, suggesting a rather higher level of genetic diversity. Analysis of population structure based on Bayesian method obtained the samilar result as the phylogeny neighbor-joining (NJ) method. The results indicated that the whole set of 143 core landraces could be clustered into two distinct groups. All landraces from Guangdong, Hainan, and 15 landraces from Jiangxi were clustered into group 1, while those from the other regions of SR formed the group 2. The results from the analysis of genetic diversity showed that both of groups possessed a similar gene diversity, but group 1 possessed relatively lower mean alleles per locus (6.63) and distinct alleles (91) than group 2 (7.94 and 110, respectively). The relatively high richness of total alleles and distinct alleles preserved in the core landraces from SR suggested that all these germplasm could be useful resources in germplasm enhancement and maize breeding in China.Key words :maize, core landraces, genetic diversity, population structureINTRODUCTIONMaize has been grown in China for nearly 500 years since its first introduction into this second biggest pro-duction country in the world. Currently, there are six different maize growing regions throughout the coun-try according to the ecological conditions and farming systems, including three major production regions,i.e., the North Spring Maize Region, the Huang-Huai-Hai Summer Maize Region, and the Southwest MaizeRegion, and three minor regions, i.e., the South Maize Region, the Northwest Maize Region, and the Qingzang Plateau Maize Region. The South Maize Region (SR)is specific because of its importance in origin of Chi-nese maize. It is hypothesized that Chinese maize is introduced mainly from two routes. One is called the land way in which maize was first brought to Tibet from India, then to Sichuan Province in southwestern China. The other way is that maize dispersed via the oceans, first shipped to the coastal areas of southeast China by boats, and then spread all round the country1252LIU Zhi-zhai et al.(Xu 2001; Zhou 2000). SR contains all of the coastal provinces and regions lie in southeastern China.In the long-term cultivation history of maize in south-ern China, numerous landraces have been formed, in which a great amount of genetic variation was observed (Li 1998). Similar to the hybrid swapping in Europe (Reif et al. 2005a), the maize landraces have been al-most replaced by hybrids since the 1950s in China (Li 1998). However, some landraces with good adapta-tions and yield performances are still grown in a few mountainous areas of this region (Liu et al.1999). Through a great effort of collection since the 1950s, 13521 accessions of maize landraces have been cur-rently preserved in China National Genebank (CNG), and a core collection of these landraces was established (Li et al. 2004). In this core collection, a total of 143 maize landrace accessions were collected from the South Maize Region (SR) (Table 1).Since simple sequence repeat ( SSR ) markers were firstly used in human genetics (Litt and Luty 1989), it now has become one of the most widely used markers in the related researches in crops (Melchinger et al. 1998; Enoki et al. 2005), especially in the molecular characterization of genetic resources, e.g., soybean [Glycine max (L.) Merr] (Xie et al. 2005), rice (Orya sativa L.) (Garris et al. 2005), and wheat (Triticum aestivum) (Chao et al. 2007). In maize (Zea mays L.), numerous studies focusing on the genetic diversity and population structure of landraces and inbred lines in many countries and regions worldwide have been pub-lished (Liu et al. 2003; Vegouroux et al. 2005; Reif et al. 2006; Wang et al. 2008). These activities of documenting genetic diversity and population structure of maize genetic resources have facilitated the under-standing of genetic bases of maize landraces, the utili-zation of these resources, and the mining of favorable alleles from landraces. Although some studies on ge-netic diversity of Chinese maize inbred lines were con-ducted (Yu et al. 2007; Wang et al. 2008), the general profile of genetic diversity in Chinese maize landraces is scarce. Especially, there are not any reports on ge-netic diversity of the maize landraces collected from SR, a possibly earliest maize growing area in China. In this paper, a total of 143 landraces from SR listed in the core collection of CNG were genotyped by using SSR markers, with the aim of revealing genetic diver-sity of the landraces from SR (Table 2) of China and examining genetic relationships and population struc-ture of these landraces.MATERIALS AND METHODSPlant materials and DNA extractionTotally, 143 landraces from SR which are listed in the core collection of CNG established by sequential strati-fication method (Liu et al. 2004) were used in the present study. Detailed information of all these landrace accessions is listed in Table 1. For each landrace, DNA sample was extracted by a CTAB method (Saghi-Maroof et al. 1984) from a bulk pool constructed by an equal-amount of leaves materials sampled from 15 random-chosen plants of each landrace according to the proce-dure of Reif et al. (2005b).SSR genotypingA total of 54 simple sequence repeat (SSR) markers covering the entire maize genome were screened to fin-gerprint all of the 143 core landrace accessions (Table 3). 5´ end of the left primer of each locus was tailed by an M13 sequence of 5´-CACGACGTTGTAAAACGAC-3´. PCR amplification was performed in a 15 L reac-tion containing 80 ng of template DNA, 7.5 mmol L-1 of each of the four dNTPs, 1×Taq polymerase buffer, 1.5 mmol L-1 MgCl2, 1 U Taq polymerase (Tiangen Biotech Co. Ltd., Beijing, China), 1.2 mol L-1 of forward primer and universal fluorescent labeled M13 primer, and 0.3 mol L-1 of M13 sequence tailed reverse primer (Schuelke 2000). The amplification was carried out in a 96-well DNA thermal cycler (GeneAmp PCR System 9700, Applied Biosystem, USA). PCR products were size-separated on an ABI Prism 3730XL DNA sequencer (HitachiHigh-Technologies Corporation, Tokyo, Japan) via the software packages of GENEMAPPER and GeneMarker ver. 6 (SoftGenetics, USA).Data analysesAverage number of alleles per locus and average num-ber of group-specific alleles per locus were identifiedAnalysis of Genetic Diversity and Population Structure of Maize Landraces from the South Maize Region of China 1253Table 1 The detailed information about the landraces used in the present studyPGS revealed by Structure1) NJ dendragram revealed Group 1 Group 2 by phylogenetic analysis140-150tian 00120005AnH-06Jingde Anhui 0.0060.994Group 2170tian00120006AnH-07Jingde Anhui 0.0050.995Group 2Zixihuangyumi00120007AnH-08Zixi Anhui 0.0020.998Group 2Zixibaihuangzayumi 00120008AnH-09Zixi Anhui 0.0030.997Group 2Baiyulu 00120020AnH-10Yuexi Anhui 0.0060.994Group 2Wuhuazi 00120021AnH-11Yuexi Anhui 0.0030.997Group 2Tongbai 00120035AnH-12Tongling Anhui 0.0060.994Group 2Yangyulu 00120036AnH-13Yuexi Anhui 0.0040.996Group 2Huangli 00120037AnH-14Tunxi Anhui 0.0410.959Group 2Baiyumi 00120038AnH-15Tunxi Anhui 0.0030.997Group 2Dapigu00120039AnH-16Tunxi Anhui 0.0350.965Group 2150tianbaiyumi 00120040AnH-17Xiuning Anhui 0.0020.998Group 2Xiuning60tian 00120042AnH-18Xiuning Anhui 0.0040.996Group 2Wubaogu 00120044AnH-19ShitaiAnhui 0.0020.998Group 2Kuyumi00130001FuJ-01Shanghang Fujian 0.0050.995Group 2Zhongdouyumi 00130003FuJ-02Shanghang Fujian 0.0380.962Group 2Baixinyumi 00130004FuJ-03Liancheng Fujian 0.0040.996Group 2Hongxinyumi 00130005FuJ-04Liancheng Fujian 0.0340.966Group 2Baibaogu 00130008FuJ-05Changding Fujian 0.0030.997Group 2Huangyumi 00130011FuJ-06Jiangyang Fujian 0.0020.998Group 2Huabaomi 00130013FuJ-07Shaowu Fujian 0.0020.998Group 2Huangbaomi 00130014FuJ-08Songxi Fujian 0.0020.998Group 2Huangyumi 00130016FuJ-09Wuyishan Fujian 0.0460.954Group 2Huabaogu 00130019FuJ-10Jian’ou Fujian 0.0060.994Group 2Huangyumi 00130024FuJ-11Guangze Fujian 0.0010.999Group 2Huayumi 00130025FuJ-12Nanping Fujian 0.0040.996Group 2Huangyumi 00130026FuJ-13Nanping Fujian 0.0110.989Group 2Hongbaosu 00130027FuJ-14Longyan Fujian 0.0160.984Group 2Huangfansu 00130029FuJ-15Loangyan Fujian 0.0020.998Group 2Huangbaosu 00130031FuJ-16Zhangping Fujian 0.0060.994Group 2Huangfansu 00130033FuJ-17Zhangping Fujian0.0040.996Group 2Baolieyumi 00190001GuangD-01Guangzhou Guangdong 0.9890.011Group 1Nuomibao (I)00190005GuangD-02Shixing Guangdong 0.9740.026Group 1Nuomibao (II)00190006GuangD-03Shixing Guangdong 0.9790.021Group 1Zasehuabao 00190010GuangD-04Lechang Guangdong 0.9970.003Group 1Zihongmi 00190013GuangD-05Lechang Guangdong 0.9880.012Group 1Jiufengyumi 00190015GuangD-06Lechang Guangdong 0.9950.005Group 1Huangbaosu 00190029GuangD-07MeiGuangdong 0.9970.003Group 1Bailibao 00190032GuangD-08Xingning Guangdong 0.9980.002Group 1Nuobao00190038GuangD-09Xingning Guangdong 0.9980.002Group 1Jinlanghuang 00190048GuangD-10Jiangcheng Guangdong 0.9960.004Group 1Baimizhenzhusu 00190050GuangD-11Yangdong Guangdong 0.9940.006Group 1Huangmizhenzhusu 00190052GuangD-12Yangdong Guangdong 0.9930.007Group 1Baizhenzhu 00190061GuangD-13Yangdong Guangdong 0.9970.003Group 1Baiyumi 00190066GuangD-14Wuchuan Guangdong 0.9880.012Group 1Bendibai 00190067GuangD-15Suixi Guangdong 0.9980.002Group 1Shigubaisu 00190068GuangD-16Gaozhou Guangdong 0.9960.004Group 1Zhenzhusu 00190069GuangD-17Xinyi Guangdong 0.9960.004Group 1Nianyaxixinbai 00190070GuangD-18Huazhou Guangdong 0.9960.004Group 1Huangbaosu 00190074GuangD-19Xinxing Guangdong 0.9950.005Group 1Huangmisu 00190076GuangD-20Luoding Guangdong 0.940.060Group 1Huangmi’ai 00190078GuangD-21Luoding Guangdong 0.9980.002Group 1Bayuemai 00190084GuangD-22Liannan Guangdong 0.9910.009Group 1Baiyumi 00300001HaiN-01Haikou Hainan 0.9960.004Group 1Baiyumi 00300003HaiN-02Sanya Hainan 0.9970.003Group 1Hongyumi 00300004HaiN-03Sanya Hainan 0.9980.002Group 1Baiyumi00300011HaiN-04Tongshi Hainan 0.9990.001Group 1Zhenzhuyumi 00300013HaiN-05Tongshi Hainan 0.9980.002Group 1Zhenzhuyumi 00300015HaiN-06Qiongshan Hainan 0.9960.004Group 1Aiyumi 00300016HaiN-07Qiongshan Hainan 0.9960.004Group 1Huangyumi 00300021HaiN-08Qionghai Hainan 0.9970.003Group 1Y umi 00300025HaiN-09Qionghai Hainan 0.9870.013Group 1Accession name Entry code Analyzing code Origin (county/city)Province/Region1254LIU Zhi-zhai et al .Baiyumi00300032HaiN-10Tunchang Hainan 0.9960.004Group 1Huangyumi 00300051HaiN-11Baisha Hainan 0.9980.002Group 1Baihuangyumi 00300055HaiN-12BaishaHainan 0.9970.003Group 1Machihuangyumi 00300069HaiN-13Changjiang Hainan 0.9900.010Group 1Hongyumi00300073HaiN-14Dongfang Hainan 0.9980.002Group 1Xiaohonghuayumi 00300087HaiN-15Lingshui Hainan 0.9980.002Group 1Baiyumi00300095HaiN-16Qiongzhong Hainan 0.9950.005Group 1Y umi (Baimai)00300101HaiN-17Qiongzhong Hainan 0.9980.002Group 1Y umi (Xuemai)00300103HaiN-18Qiongzhong Hainan 0.9990.001Group 1Huangmaya 00100008JiangS-10Rugao Jiangsu 0.0040.996Group 2Bainian00100012JiangS-11Rugao Jiangsu 0.0080.992Group 2Bayebaiyumi 00100016JiangS-12Rudong Jiangsu 0.0040.996Group 2Chengtuohuang 00100021JiangS-13Qidong Jiangsu 0.0050.995Group 2Xuehuanuo 00100024JiangS-14Qidong Jiangsu 0.0020.998Group 2Laobaiyumi 00100032JiangS-15Qidong Jiangsu 0.0050.995Group 2Laobaiyumi 00100033JiangS-16Qidong Jiangsu 0.0010.999Group 2Huangwuye’er 00100035JiangS-17Hai’an Jiangsu 0.0030.997Group 2Xiangchuanhuang 00100047JiangS-18Nantong Jiangsu 0.0060.994Group 2Huangyingzi 00100094JiangS-19Xinghua Jiangsu 0.0040.996Group 2Xiaojinhuang 00100096JiangS-20Yangzhou Jiangsu 0.0010.999Group 2Liushizi00100106JiangS-21Dongtai Jiangsu 0.0030.997Group 2Kangnandabaizi 00100108JiangS-22Dongtai Jiangsu 0.0020.998Group 2Shanyumi 00140020JiangX-01Dexing Jiangxi 0.9970.003Group 1Y umi00140024JiangX-02Dexing Jiangxi 0.9970.003Group 1Tianhongyumi 00140027JiangX-03Yushan Jiangxi 0.9910.009Group 1Hongganshanyumi 00140028JiangX-04Yushan Jiangxi 0.9980.002Group 1Zaoshuyumi 00140032JiangX-05Qianshan Jiangxi 0.9970.003Group 1Y umi 00140034JiangX-06Wannian Jiangxi 0.9970.003Group 1Y umi 00140038JiangX-07De’an Jiangxi 0.9940.006Group 1Y umi00140045JiangX-08Wuning Jiangxi 0.9740.026Group 1Chihongyumi 00140049JiangX-09Wanzai Jiangxi 0.9920.008Group 1Y umi 00140052JiangX-10Wanzai Jiangxi 0.9930.007Group 1Huayumi 00140060JiangX-11Jing’an Jiangxi 0.9970.003Group 1Baiyumi 00140065JiangX-12Pingxiang Jiangxi 0.9940.006Group 1Huangyumi00140066JiangX-13Pingxiang Jiangxi 0.9680.032Group 1Nuobaosuhuang 00140068JiangX-14Ruijin Jiangxi 0.9950.005Group 1Huangyumi 00140072JiangX-15Xinfeng Jiangxi 0.9960.004Group 1Wuningyumi 00140002JiangX-16Jiujiang Jiangxi 0.0590.941Group 2Tianyumi 00140005JiangX-17Shangrao Jiangxi 0.0020.998Group 2Y umi 00140006JiangX-18Shangrao Jiangxi 0.0310.969Group 2Baiyiumi 00140012JiangX-19Maoyuan Jiangxi 0.0060.994Group 260riyumi 00140016JiangX-20Maoyuan Jiangxi 0.0020.998Group 2Shanyumi 00140019JiangX-21Dexing Jiangxi 0.0050.995Group 2Laorenya 00090002ShangH-01Chongming Shanghai 0.0050.995Group 2Jinmeihuang 00090004ShangH-02Chongming Shanghai 0.0020.998Group 2Zaobaiyumi 00090006ShangH-03Chongming Shanghai 0.0020.998Group 2Chengtuohuang 00090007ShangH-04Chongming Shanghai 0.0780.922Group 2Benyumi (Huang)00090008ShangH-05Shangshi Shanghai 0.0020.998Group 2Bendiyumi 00090010ShangH-06Shangshi Shanghai 0.0040.996Group 2Baigengyumi 00090011ShangH-07Jiading Shanghai 0.0020.998Group 2Huangnuoyumi 00090012ShangH-08Jiading Shanghai 0.0040.996Group 2Huangdubaiyumi 00090013ShangH-09Jiading Shanghai 0.0440.956Group 2Bainuoyumi 00090014ShangH-10Chuansha Shanghai 0.0010.999Group 2Laorenya 00090015ShangH-11Shangshi Shanghai 0.0100.990Group 2Xiaojinhuang 00090016ShangH-12Shangshi Shanghai 0.0050.995Group 2Gengbaidayumi 00090017ShangH-13Shangshi Shanghai 0.0020.998Group 2Nongmeiyihao 00090018ShangH-14Shangshi Shanghai 0.0540.946Group 2Chuanshazinuo 00090020ShangH-15Chuansha Shanghai 0.0550.945Group 2Baoanshanyumi 00110004ZheJ-01Jiangshan Zhejiang 0.0130.987Group 2Changtaixizi 00110005ZheJ-02Jiangshan Zhejiang 0.0020.998Group 2Shanyumibaizi 00110007ZheJ-03Jiangshan Zhejiang 0.0020.998Group 2Kaihuajinyinbao 00110017ZheJ-04Kaihua Zhejiang 0.0100.990Group 2Table 1 (Continued from the preceding page)PGS revealed by Structure 1) NJ dendragram revealed Group1 Group2 by phylogenetic analysisAccession name Entry code Analyzing code Origin (county/city)Province/RegoinAnalysis of Genetic Diversity and Population Structure of Maize Landraces from the South Maize Region of China 1255Liputianzi00110038ZheJ-05Jinhua Zhejiang 0.0020.998Group 2Jinhuaqiuyumi 00110040ZheJ-06Jinhua Zhejiang 0.0050.995Group 2Pujiang80ri 00110069ZheJ-07Pujiang Zhejiang 0.0210.979Group 2Dalihuang 00110076ZheJ-08Yongkang Zhejiang 0.0140.986Group 2Ziyumi00110077ZheJ-09Yongkang Zhejiang 0.0020.998Group 2Baiyanhandipinzhong 00110078ZheJ-10Yongkang Zhejiang 0.0030.997Group 2Duosuiyumi00110081ZheJ-11Wuyi Zhejiang 0.0020.998Group 2Chun’an80huang 00110084ZheJ-12Chun’an Zhejiang 0.0020.998Group 2120ribaiyumi 00110090ZheJ-13Chun’an Zhejiang 0.0020.998Group 2Lin’anliugu 00110111ZheJ-14Lin’an Zhejiang 0.0030.997Group 2Qianhuangyumi00110114ZheJ-15Lin’an Zhejiang 0.0030.997Group 2Fenshuishuitianyumi 00110118ZheJ-16Tonglu Zhejiang 0.0410.959Group 2Kuihualiugu 00110119ZheJ-17Tonglu Zhejiang 0.0030.997Group 2Danbaihuang 00110122ZheJ-18Tonglu Zhejiang 0.0020.998Group 2Hongxinma 00110124ZheJ-19Jiande Zhejiang 0.0030.997Group 2Shanyumi 00110136ZheJ-20Suichang Zhejiang 0.0030.997Group 2Bai60ri 00110143ZheJ-21Lishui Zhejiang 0.0050.995Group 2Zeibutou 00110195ZheJ-22Xianju Zhejiang 0.0020.998Group 2Kelilao00110197ZheJ-23Pan’an Zhejiang 0.0600.940Group 21)The figures refered to the proportion of membership that each landrace possessed.Table 1 (Continued from the preceding page)PGS revealed by Structure 1) NJ dendragram revealed Group 1 Group 2 by phylogenetic analysisAccession name Entry code Analyzing code Origin (county/city)Province/Regoin Table 2 Construction of two phylogenetic groups (SSR-clustered groups) and their correlation with geographical locationsGeographical location SSR-clustered groupChi-square testGroup 1Group 2Total Guangdong 2222 χ2 = 124.89Hainan 1818P < 0.0001Jiangxi 15621Anhui 1414Fujian 1717Jiangsu 1313Shanghai 1515Zhejiang 2323Total5588143by the software of Excel MicroSatellite toolkit (Park 2001). Average number of alleles per locus was calcu-lated by the formula rAA rj j¦1, with the standarddeviation of1)()(12¦ r A AA rj jV , where A j was thenumber of distinct alleles at locus j , and r was the num-ber of loci (Park 2001).Unbiased gene diversity also known as expected heterozygosity, observed heterozygosity for each lo-cus and average gene diversity across the 54 SSR loci,as well as model-based groupings inferred by Struc-ture ver. 2.2, were calculated by the softwarePowerMarker ver.3.25 (Liu et al . 2005). Unbiased gene diversity for each locus was calculated by˅˄¦ 2ˆ1122ˆi x n n h , where 2ˆˆ2ˆ2¦¦z ji ijij i X X x ,and ij X ˆwas the frequency of genotype A i A jin the sample, and n was the number of individuals sampled.The average gene diversity across 54 loci was cal-culated as described by Nei (1987) as follows:rh H rj j ¦1ˆ, with the variance ,whereThe average observed heterozygosity across the en-tire loci was calculated as described by (Hedrick 1983)as follows: r jrj obsobs n h h ¦1, with the standard deviationrn h obs obsobs 1V1256LIU Zhi-zhai et al.Phylogenetic analysis and population genetic structureRelationships among all of the 143 accessions collected from SR were evaluated by using the unweighted pair group method with neighbor-joining (NJ) based on the log transformation of the proportion of shared alleles distance (InSPAD) via PowerMarker ver. 3.25 (FukunagaTable 3 The PIC of each locus and the number of alleles detected by 54 SSRsLocus Bin Repeat motif PIC No. of alleles Description 2)bnlg1007y51) 1.02AG0.7815Probe siteumc1122 1.06GGT0.639Probe siteumc1147y41) 1.07CA0.2615Probe sitephi961001) 2.00ACCT0.298Probe siteumc1185 2.03GC0.7215ole1 (oleosin 1)phi127 2.08AGAC0.577Probe siteumc1736y21) 2.09GCA T0.677Probe sitephi453121 3.01ACC0.7111Probe sitephi374118 3.03ACC0.477Probe sitephi053k21) 3.05A TAC0.7910Probe sitenc004 4.03AG0.4812adh2 (alcohol dehydrogenase 2)bnlg490y41) 4.04T A0.5217Probe sitephi079 4.05AGATG0.495gpc1(glyceraldehyde-3-phosphate dehydrogenase 1) bnlg1784 4.07AG0.6210Probe siteumc1574 4.09GCC0.719sbp2 (SBP-domain protein 2)umc1940y51) 4.09GCA0.4713Probe siteumc1050 4.11AA T0.7810cat3 (catalase 3)nc130 5.00AGC0.5610Probe siteumc2112y31) 5.02GA0.7014Probe sitephi109188 5.03AAAG0.719Probe siteumc1860 5.04A T0.325Probe sitephi085 5.07AACGC0.537gln4 (glutamine synthetase 4)phi331888 5.07AAG0.5811Probe siteumc1153 5.09TCA0.7310Probe sitephi075 6.00CT0.758fdx1 (ferredoxin 1)bnlg249k21) 6.01AG0.7314Probe sitephi389203 6.03AGC0.416Probe sitephi299852y21) 6.07AGC0.7112Probe siteumc1545y21)7.00AAGA0.7610hsp3(heat shock protein 3)phi1127.01AG0.5310o2 (opaque endosperm 2)phi4207018.00CCG0.469Probe siteumc13598.00TC0.7814Probe siteumc11398.01GAC0.479Probe siteumc13048.02TCGA0.335Probe sitephi1158.03A TAC0.465act1(actin1)umc22128.05ACG0.455Probe siteumc11218.05AGAT0.484Probe sitephi0808.08AGGAG0.646gst1 (glutathione-S-transferase 1)phi233376y11)8.09CCG0.598Probe sitebnlg12729.00AG0.8922Probe siteumc20849.01CTAG0.498Probe sitebnlg1520k11)9.01AG0.5913Probe sitephi0659.03CACCT0.519pep1(phosphoenolpyruvate carboxylase 1)umc1492y131)9.04GCT0.2514Probe siteumc1231k41)9.05GA0.2210Probe sitephi1084119.06AGCT0.495Probe sitephi4488809.06AAG0.7610Probe siteumc16759.07CGCC0.677Probe sitephi041y61)10.00AGCC0.417Probe siteumc1432y61)10.02AG0.7512Probe siteumc136710.03CGA0.6410Probe siteumc201610.03ACAT0.517pao1 (polyamine oxidase 1)phi06210.04ACG0.337mgs1 (male-gametophyte specific 1)phi07110.04GGA0.515hsp90 (heat shock protein, 90 kDa)1) These primers were provided by Beijing Academy of Agricultural and Forestry Sciences (Beijing, China).2) Searched from Analysis of Genetic Diversity and Population Structure of Maize Landraces from the South Maize Region of China1257et al. 2005). The unrooted phylogenetic tree was finally schematized with the software MEGA (molecular evolu-tionary genetics analysis) ver. 3.1 (Kumar et al. 2004). Additionally, a chi-square test was used to reveal the correlation between the geographical origins and SSR-clustered groups through FREQ procedure implemented in SAS ver. 9.0 (2002, SAS Institute, Inc.).In order to reveal the population genetic structure (PGS) of 143 landrace accessions, a Bayesian approach was firstly applied to determine the number of groups (K) that these materials should be assigned by the soft-ware BAPS (Bayesian Analysis of Population Structure) ver.5.1. By using BAPS, a fixed-K clustering proce-dure was applied, and with each separate K, the num-ber of runs was set to 100, and the value of log (mL) was averaged to determine the appropriate K value (Corander et al. 2003; Corander and Tang 2007). Since the number of groups were determined, a model-based clustering analysis was used to assign all of the acces-sions into the corresponding groups by an admixture model and a correlated allele frequency via software Structure ver.2.2 (Pritchard et al. 2000; Falush et al. 2007), and for the given K value determined by BAPS, three independent runs were carried out by setting both the burn-in period and replication number 100000. The threshold probability assigned individuals into groupswas set by 0.8 (Liu et al. 2003). The PGS result carried out by Structure was visualized via Distruct program ver. 1.1 (Rosenberg 2004).RESULTSGenetic diversityA total of 517 alleles were detected by the whole set of54 SSRs covering the entire maize genome through all of the 143 maize landraces, with an average of 9.57 alleles per locus and ranged from 4 (umc1121) to 22 (bnlg1272) (Table 3). Among all the alleles detected, the number of distinct alleles accounted for 132 (25.53%), with an av-erage of 2.44 alleles per locus. The distinct alleles dif-fered significantly among the landraces from different provinces/regions, and the landraces from Guangdong, Fujian, Zhejiang, and Shanghai possessed more distinct alleles than those from the other provinces/regions, while those from southern Anhui possessed the lowest distinct alleles, only counting for 3.28% of the total (Table 4).Table 4 The genetic diversity within eight provinces/regions and groups revealed by 54 SSRsProvince/Region Sample size Allele no.1)Distinct allele no.Gene diversity (expected heterozygosity)Observed heterozygosity Anhui14 4.28 (4.19) 69 (72.4)0.51 (0.54)0.58 (0.58)Fujian17 4.93 (4.58 80 (79.3)0.56 (0.60)0.63 (0.62)Guangdong22 5.48 (4.67) 88 (80.4)0.57 (0.59)0.59 (0.58)Hainan18 4.65 (4.26) 79 (75.9)0.53 (0.57)0.55 (0.59)Jiangsu13 4.24 700.500.55Jiangxi21 4.96 (4.35) 72 (68.7)0.56 (0.60)0.68 (0.68)Shanghai15 5.07 (4.89) 90 (91.4)0.55 (0.60)0.55 (0.55)Zhejiang23 5.04 (4.24) 85 (74)0.53 (0.550.60 (0.61)Total/average1439.571320.610.60GroupGroup 155 6.63 (6.40) 91 (89.5)0.57 (0.58)0.62 (0.62)Group 2887.94 (6.72)110 (104.3)0.57 (0.57)0.59 (0.58)Total/Average1439.571320.610.60Provinces/Regions within a groupGroup 1Total55 6.69 (6.40) 910.57 (0.58)0.62 (0.62)Guangdong22 5.48 (4.99) 86 (90.1)0.57 (0.60)0.59 (0.58)Hainan18 4.65 (4.38) 79 (73.9)0.53 (0.56)0.55 (0.59)Jiangxi15 4.30 680.540.69Group 2Total887.97 (6.72)110 (104.3)0.57 (0.57)0.59 (0.58)Anhui14 4.28 (3.22) 69 (63.2)0.51 (0.54)0.58 (0.57)Fujian17 4.93 (3.58) 78 (76.6)0.56 (0.60)0.63 (0.61)Jiangsu13 4.24 (3.22) 71 (64.3)0.50 (0.54)0.55 (0.54)Jiangxi6 3.07 520.460.65Shanghai15 5.07 (3.20) 91 (84.1)0.55 (0.60)0.55 (0.54)Zhejiang23 5.04 (3.20) 83 (61.7)0.53 (0.54)0.60 (0.58)1258LIU Zhi-zhai et al.Among the 54 loci used in the study, 16 (or 29.63%) were dinucleotide repeat SSRs, which were defined as type class I-I, the other 38 loci were SSRs with a longer repeat motifs, and two with unknown repeat motifs, all these 38 loci were defined as the class of I-II. In addition, 15 were located within certain functional genes (defined as class II-I) and the rest were defined as class II-II. The results of comparison indicated that the av-erage number of alleles per locus captured by class I-I and II-II were 12.88 and 10.05, respectively, which were significantly higher than that by type I-II and II-I (8.18 and 8.38, respectively). The gene diversity re-vealed by class I-I (0.63) and II-I (0.63) were some-what higher than by class I-II (0.60) and II-II (0.60) (Table 5).Genetic relationships of the core landraces Overall, 143 landraces were clustered into two groups by using neighbor-joining (NJ) method based on InSPAD. All the landraces from provinces of Guangdong and Hainan and 15 of 21 from Jiangxi were clustered together to form group 1, and the other 88 landraces from the other provinces/regions formed group 2 (Fig.-B). The geographical origins of all these 143 landraces with the clustering results were schematized in Fig.-D. Revealed by the chi-square test, the phylogenetic results (SSR-clustered groups) of all the 143 landraces from provinces/regions showed a significant correlation with their geographical origin (χ2=124.89, P<0.0001, Table 2).Revealed by the phylogenetic analysis based on the InSPAD, the minimum distance was observed as 0.1671 between two landraces, i.e., Tianhongyumi (JiangX-03) and Hongganshanyumi (JiangX-04) collected from Jiangxi Province, and the maximum was between two landraces of Huangbaosu (FuJ-16) and Hongyumi (HaiN-14) collected from provinces of Fujian and Hainan, respectively, with the distance of 1.3863 (data not shown). Two landraces (JiangX-01 and JiangX-21) collected from the same location of Dexing County (Table 1) possessing the same names as Shanyumi were separated to different groups, i.e., JiangX-01 to group1, while JiangX-21 to group 2 (Table 1). Besides, JiangX-01 and JiangX-21 showed a rather distant distance of 0.9808 (data not shown). These results indicated that JiangX-01 and JiangX-21 possibly had different ances-tral origins.Population structureA Bayesian method was used to detect the number of groups (K value) of the whole set of landraces from SR with a fixed-K clustering procedure implemented in BAPS software ver. 5.1. The result showed that all of the 143 landraces could also be assigned into two groups (Fig.-A). Then, a model-based clustering method was applied to carry out the PGS of all the landraces via Structure ver. 2.2 by setting K=2. This method as-signed individuals to groups based on the membership probability, thus the threshold probability 0.80 was set for the individuals’ assignment (Liu et al. 2003). Accordingly, all of the 143 landraces were divided into two distinct model-based groups (Fig.-C). The landraces from Guangdong, Hainan, and 15 landraces from Jiangxi formed one group, while the rest 6 landraces from the marginal countries of northern Jiangxi and those from the other provinces formed an-other group (Table 1, Fig.-D). The PGS revealed by the model-based approach via Structure was perfectly consistent with the relationships resulted from the phy-logenetic analysis via PowerMarker (Table 1).DISCUSSIONThe SR includes eight provinces, i.e., southern Jiangsu and Anhui, Shanghai, Zhejiang, Fujian, Jiangxi, Guangdong, and Hainan (Fig.-C), with the annual maize growing area of about 1 million ha (less than 5% of theTable 5 The genetic diversity detected with different types of SSR markersType of locus No. of alleles Gene diversity Expected heterozygosity PIC Class I-I12.880.630.650.60 Class I-II8.180.600.580.55 Class II-I8.330.630.630.58。
四组细胞瘤块切片进行免疫组化分析...
图ｉ克隆的鉴定Ａ：ＲＴ－－ＰＣＲ鉴定结果Ｂ：ｗｅｓｔｅｒｎｂｌｏｔ鉴定结果Ｉ：阳性对照办２：ＭＤＡ－－ＭＢ－－４３５／Ｉｋｂｌ（Ｌ），３：ＭＤＡ一船一４３５几ＫＢｌ（Ｈ）。
４：ＭＤＡ一蛐－－４３５２，５：ＭＤＡ一船－－４３５／ｖｅｃ图２各组细胞ｔｒａｎｓｗｅｌｌ细胞侵袭实验数据为４株肿瘤细胞３次独立实验的均数±标准差。
误差线代表标准误。
２图３各细胞ＭＭＰ一２、删Ｐ一９、ＶＥＧＦ、ＢＦＧＦｍＲＮＡ的表达隋况Ａ：ＲＴ—ＰＣＲ检测结果ｌ、２、３，４分别代表为ＭＤＡ－ＭＢ－４３５细胞，ＭＤＡ一蛐一４３５／ｖｅｃ细胞－叭—姗一４３５／ＬＩ（Ｂ１（Ｈ）细胞、Ｄ为ＭＤＡ－ＭＢ－４３５／ＬＫＢｌ（Ｌ）细胞３图５ＭＭＰ一２，删Ｐ一９活性的测定Ａ：凝胶酶谱ＭＭＰ－２，姗Ｐ一９检测结果１、２、３．４分别代表为ＭＤＡ－惦一４３５细胞、ＭＩ）Ａ－ＭＢ一４３５／ｖｅｃ细胞ＭＩ）Ａ—ＭＢ－４３５／ＬＫＢｌ（Ｈ）细胞，Ｄ为～ｆ１）Ａ－１４Ｂ一４３５／ＬＫＢｌ（Ｌ）细胞图４各细胞ＭＭＰ一２、姗Ｐ一９、ＶＥＧＦ、ＢＦＧＦ蛋白的表达情况Ａ：ｗｅｓｔｅｒｎｂｌｏｔ检测结果ｌ、２，３，４分别代表为ＭＤＡ－ＭＢ－４３５细胞、ｌ￥）Ａ－ＭＢ－４３５／ｖｅｃ细胞盼Ａ一邺一４３５几Ｉ（Ｂｌｍ）细胞、Ｄ为如Ａ－船一４３５／ＬＫＢｌ（Ｌ）细胞图６人乳癌裸鼠原位移植瘤情况ＭＤＡ·ＭＢ－４３５／ＬＫＢｌ（Ｌ）ＭＤＡ·ＭＢ·４３５／ＬＫＢｌ（Ｈ）ＭＤＡ—ＭＢ一４３５ＮＥＣＭＤＡ－ＭＢ－４３５图７各组鼠肺转移灶比较ＰｕＩｍ。
ｎａｒｙｍｅｌａｓｔａｓｌｓｔｕｍ。
阽（ＨＥ’１００）图８各移植瘤ＭＭＰ一２、ＭＭＰ－－９、ＶＥＧＦ、ＢＦＧＦ蛋白的表达情况Ａ：ｗｅｓｔｅｒｎｂｌｏｔ检测结果１、２、３代表ＭＤＡ－ＭＢ－４３５细胞、４，５，６代表ｄ４ＤＡ－ＭＢ－４３５／ｖｅｃ细胞７，８，９代表ＭＤＡ－ＭＢ－４３５／ＬＫＢｌ（Ｌ）细胞、１０，１１．１２代表ｈⅢＡ—ｍ一４３５／ｕ（Ｂｌ（Ｈ）细胞图９各组瘤组织微血管密度变化图１０ＬＫＢｌ基因与微血管密度变化的关系LKB1基因与人乳腺癌细胞生长和侵袭的相关性研究作者：庄志刚学位授予单位：复旦大学1.Westerman AM.Entius MM.de Baar E Peutz-Jeghers syndrome:78-year follow-up of the original family 19992.Tiainen M.Ylikorkala A.Makela TP Growth arrest by the LKB1 tumor suppressor:induction ofp21(WAFl/CIP1) 2002(13)3.Shen Z.Wen n F The tumor suppressor gene LKB1 is assoclate with prognosis in human breast carcinoma 2002(07)4.Sapkota GP.Boudeau J.Deak M Phosphorylation of the protein kinase mutated in Peutz-Jeghers cancer syndrome,LKB1/STK11,at Ser431 by p90(RSK)and cAMP-dependent protein kinase,but not its farnesylation at Cys(433),is essential for LKB1 to suppress cell vrowth 2001(22)5.8apkota GP.Boudeau J.Deak M Identification and characterization of four novel phosphorylationsites(Ser31,Ser325,Thr336 and Thr366)on LKB1/STK11,the protein kinase mutated in Peutz-Jeghers cancer syndrome 2002(02)6.Forster LF.Defres S.Goudie DR An investigation of the Peutz-Jeghers gene(LKB1)in sporadic breast and colon cancers 20007.Chen J Lindblom Germline mutation screening of the STK11/LKB1 gene in familial breast cancer with LOH on 19p 20008.Kleiner DE.Stetler-Stevenson WG Matrix metalloproteinases and metastasis 19999.McCawley LJ.Matrisian LM Matrix metalloproteinases:they are not just for matrix anymore! 200110.Hojilla CV.Mohammed FF.Khokha R Matrix metalloproteinases and theirtissue inhibitors direct cell fate during cancer development 2003(10)11.Stamenkovic I Matrix metalloproteinases in tumor invasion andmetastasis 2000(06)12.John A.Tuszynski G The role of matrix metalloproteinases in tumor angiogenesis and tumor metastasis 2001(01)13.Freije JM.Balbin M.Pendas AM Matrix metalloproteinases and tumor progression 200314.Jones JL.Shaw JA.Pringle JH Primary breast myoepithelial cells exert an invasion-suppressor effect on breast cancer celis via paracrine down-regulation of MMP expression in fibroblasts and tumour cells 2003(04)15.Karuman P.Gozani O.Odze RD The Peutz-Jegher gene product LKB1 is a mediator of p53-dependent cell death[外文期刊] 200116.Wang JL.Sun Y.Wu S Gamma-irradiation induces matrix metalloproteinase Ⅱ expression in a p53-dependent manner 2000(04)17.Folkmen J Tumor angiogenesis:Therapeutic implication 197121.Tsutsui S.Kume M.Era S Prognostic value of microvessel density in invasive ductal carcinoma of the breast 2003(04)22.Weidner N.Semple J P.Welch W R.Folkman J Tumor angiogenesis and metastasis-correlat ion in invasive breast carcinoma 199123.Guidi AJ.Schnitt SJ.Fischer L Vascular permeability factor(vascular endothelial growthfactor)expresslon and anglogenesis in patients with ductal carcinoma in situ of the breast 199724.Ramanathan M.Giladi A.Leibovich SJ Regularion of vascularendothelial growth factor gene expression in murine macrophages by nitric oxide and hypoxia 2003(06)25.Ylikorkala A.Rossi DJ.Korsisaari N Vascular abnormalities and deregulation of VEGF in LKB1-deficient mice[外文期刊] 200126.Westerman AM.Entius MM.de Baar E Peutz-Jeghers syndrome:78-year follow-up of the original family 199927.Tiainen M.Ylikorkala A.Makela TP Growth arrest by the LKB1 tumor suppressor:induction ofp21(WAF1/CIP1) 2002(13)28.Jimenez AI.Fernandez P.Dominguez O Growth and molecular profile of lung cancer cells expressing ectopic LKB1:down-regulation of the phosphatidylinositol 3-phosphate kinase/PTEN pathway 2003(06)29.Ylikorkala A.Rossi DJ.Korsisaari N Vascular abnormalities and deregulation of VEGF in Lkb1-deficient mice[外文期刊] 200130.Miyoshi H.Nakau M.Ishikawa TO Gastrointestinal hamartomatous polyposis in lkb1 heterozygous knockout mice 2002(08)31.Rossi DJ.Ylikorkala A.Korsisaari N Induction of cyclooxygenase-2 in a mouse model of Peutz-Jeghers polyposis 2002(19)32.Bardeesy N.Sinha M.Hezel AF Loss of the Lkb1 tumor suppressor provokes intestinal polyposis but resistance to transformation 2002(6903)33.Nakau M.Miyoshi H.Seldin MF Hepatocelhlar carcinoma caused by loss of heterozygosity in lkb1 gene knockout mice 2002(16)34.Watts JL.Morton DG.Bestman j The C.elegans par-4 gene encodes a putative serine-threonine kinase required for establishing embryonic asymmetry35.Martin SG.St Johnston D A role for Drosophila LKB1 in anterior-posterior axis formation and epithelial polarity[外文期刊] 2003(6921)36.Baas AF.Boufeau J.Sapkota GP Activation of the tumour suppressor kinase LKB1 by the STE20-like pseudokinase STRAD 2003(12)37.Sapkota GP.Boudeau J.Deak M Phosphorylation of the protein kinase mutated in Peutz-Jeghers cancer syndrome,LKB1/STK11,at Ser431 by p90(RSK)and cAMP-dependent protein kinase,but not its farnesylation at Cys(433),is essential for LKB1 to suppress cell vrowth 2001(22)38.Sapkota GP.Boudeau J.Deak M Identification and characterization of four novel phosphorylation39.Smith DP.Rayter SI.Niederlander C LIP1,a cytoplasmic proteinfunctionally linked to the Peutz-Jeghers syndrome kinase LKB1 2001(25)40.Marignani PA.Kanai F.Carpenter CL LKB1 associates with Brg1 and is necessary for Brg1-induced growth arrest 2001(35)41.Karuman P.Gozani O.Odze RD The Peutz-Jegher gene product LKB1 is a mediator of p53-dependent cell death[外文期刊] 2001(06)42.Bignell GR.Barfoot R.Seal S Low frequency of somatic mutations in the LKB1/Peutz-Jeghers syndrome gene in sporadic breast cancer 1998(07)43.Forster LF.Defres S.Goudie DR An investigation of the Peutz-Jeghers gene(LKB1)in sporadic breast and colon cancers 2000(10)44.Chen J Lindblom Germline mutation screening of the STK11/LKB1 gene in familial breast cancer with LOH on 19p 2000(05)45.Shen Z.Wen n F The tumor suppressor gene Lkb1 is associate with prognosis in human breast carcinoma 2002(07)1.费菲原肌球蛋白-4在人类乳腺癌高低转移细胞株中的差异性表达及临床意义[学位论文]20052.沈赞抑癌基因LKB1在乳腺癌中的作用研究[学位论文]20023.王振军.严仲瑜.毕郭龙国人黑斑息肉病LKB1基因胚系突变的检测[期刊论文]-中华外科杂志2000,38 (2)4.董慧明上皮钙黏蛋白对人炎性乳腺癌细胞系生物学特性的影响[学位论文]20055.丁锦华微浸润在导管原位癌中的临床意义及MMP-2、Tenascin-C在导管原位癌中的表达[学位论文]20056.刘刚乳腺癌血管、淋巴管生成与转移预后的研究[学位论文]20037.张杰乳腺癌前哨淋巴结活检及骨髓播散肿瘤细胞的检测[学位论文]20058.王劲松Rab27A对人乳腺癌细胞生物学特性的影响及其机制的研究[学位论文]20079.宋科瑛耐药乳腺癌细胞MDR-MCF-7侵袭力增强机制探讨[学位论文]200310.李鹤成ERα和Her-2受体在人乳腺癌细胞株作用通路的实验研究及基质金属蛋白酶在淋巴结阴性乳腺癌的预后意义[学位论文]2005本文链接：/Thesis_Y952108.aspx。
Mettler Toledo 试验用品说明书
M351Glycerin, M = 92.10, z = 1Potassium hydrogen phthalate (KHP)Sodium(meta)periodate (NaIO 4)Potassium iodide, 0.6 mol/LSodium thiosulfate c =01mol/L Sodium hydroxide (NaOH)c(NaOH) = 0.1 mol/LT50, T70 and T90 Excellence Titrators1) Suitability test (Redox):IO 4-+7I -+6H ++H 2O = 4I 2+2OH -+3H 2O 4I 2+8S 2O 32-= 4S 4O 62-+ 8I -, IO 4-: z = 82) Acid base titration:HCOOH + OH -= H2O + HCOO-Titration beakers ME-101974OKI B4250 Printer(parallel cable ME-51 108 780)METTLER TOLEDONeutralization before final disposalApprox. 500 mg Glycerin (anhydrous,> 98%)DM140-SC (Redox titration)DG111-SC (Acid base titration)1.1): 60 g of sodium(meta)periodate are dissolved in a 1 L flask with 12% v/v sulfuric acid. The solution is stored in a brown titration glass bottle.1.2):- 10 mL of sodium periodate solution are pipetted into a 250mL flask and filled up with water.- 500 mg of glycerin are dissolved in 50 mL water and 50 mL of the diluted periodate solution was added. Wait for 30 min.- Blank: 50 mL of the diluted periodate solution was added to 50 mL water. Wait for 30 min.- 20 mL aliquots of both solutions were acidified with 1 mL of 1.0 mol/L HCl. 10 ml 0.6 mol/L KI-solution are added. Deion.water was added to achieve a volume of 50 mL. The released iodine was titrated with 0.1 mol/L sodium thiosulfate.2): 500 mg glycerin are weighed in a titration beaker and 50 mL deion. water were added. For a blank determination 50 mL deionized water were added separately into a titration beaker. The pH was adjusted to 7.8 for both solutions. 50 mL of the sodium periodate solution was pipetted into each beaker and covered wih a watch glass. Both solutions were allowed to stand for 30 min at room temperature. 10 mL of a PEG (polyethyleneglycol)/water mixture (1:1, v:v) were added to both solutions and allowed to stand for 20 min.1): According to USP26 the ratio between the volume of 0.1 mol/L sodium thiosulfate required for the glycerin containing mixture to that for the blank should be between0.750 and 0.765. The obtained ratio was 0.750. This ratio was regarded as a proof that the periodate solution is suitable for the analysis of the glycerin content.2) Acid base titration: The nominal concentration of the glycerin solution was 0.04995 mol/L (0.5061 g in 110 mL). This corresponds to a recovery of 98.90% (see results: mean value of the obtained glycerin concentration). Blank and sample determinations were performed in the same EQP-titration mode and the obtained blank value (mmol) was substracted from the sample value (mmol) in order to account for the acidity in the solvent not corresponding to the released formic acid.3) Oxidation of glycerin with periodate:CH 2OH-CHOH-CH 2OH + 2IO 4-= HCOOH + 2 HCHO + 2 IO 3-+ H 2OOne mole of glycerin corresponds therefore to one mole of formicacid. Formic acid (M = 46.025, z = 1) was titrated with sodium hydroxide.Literature:"USP 26 -Official monographs / Glycerin" p. 867R1=Q-B[Glycerin]; mmol R2=(Q-B[Glycerin])*C/m C=1/z ; mol/LB[Glycerin]: Blank valueThe glycerin content is determined according to USP 26 with acid base titration: Formic acid that is released from glycerin by quantitative reaction with sodium periodate is titrated with sodium hydroxide. Beforehand the suitability of the periodate solution is validated by redox titration using sodium thiosulfate as titrant.Potassium hydrogen phthalate, 80 mg Thomas HitzName: Thomas Hitz, ID Glycerin ContentRx Result Unit Name1/5 -- 27.03.2007 11:30:27R1 =0.98657 mmol ContentR2 =0.04933 mol/L Concentration 2/5 -- 27.03.2007 11:34:16R1 =0.98932 mmol ContentR2 =0.04947 mol/L Concentration 3/5 -- 27.03.2007 11:41:09R1 =0.98976 mmol ContentR2 =0.04949 mol/L Concentration 4/5 -- 27.03.2007 11:47:34R1 =0.98991 mmol ContentR2 =0.04950 mol/L Concentration 5/5 -- 27.03.2007 11:53:14R1 =0.98406 mmol ContentR2 =0.04920 mol/L Concentration StatisticsRx Name n Mean Unit s srel [%]R1 Cont.50.98792 mmol0.00255 0.258R2 Conc.50.04940 mol/L0.00013 0.264。
硅烷偶联剂改性酚醛树脂的合成
CHINA SYNTHETIC RESIN AND PLASTICS 研究与开发合 成 树 脂 及 塑 料 ， 2017， 34（6）： 17酚醛树脂是酚类与醛类在酸性或碱性催化剂作用下形成树脂的统称，是工业化最早的合成高分子材料，具有优异的黏接强度、耐水、耐热、耐磨、耐化学药品腐蚀性及化学稳定性等特点，特别是耐沸水性能最佳。
目前，酚醛树脂仍是相当重要的合成高分子材料，特别在生产耐水、耐候性木制品等，具有十分特别的意义。
酚醛树脂同样有着一些缺点，颜色太深、脆性易裂等，所以在应用上有着一定的限制［1-4］。
许多科研工作者从分子结构、聚合工艺以及共混等方面对其进行了研究，取得了一定的成效［5-9］。
由于传统的酚醛树脂在耐热性能和韧性等方面存在缺陷，在很大程度上限制了其进一步应用，而有机硅树脂具有良好的耐热性能和韧性。
本工作针对普通酚醛树脂的脆性和耐热性能的不足，采用硅烷偶联剂KH560改性酚醛树脂，在结构中引入Si—O—Si和环氧基，通过优化实验条件，对树脂的性能进行分析，期望改善树脂的耐热性能和韧性，为酚醛树脂的改性提供一种可供选择的参考方法，拓展酚醛树脂的应用领域，对推动酚醛树脂产业发展具有重要的意义。
硅烷偶联剂改性酚醛树脂的合成游胜勇1，戴润英2*，董晓娜1，李 玲1，陈衍华1，曹 修1（1.江西省科学院应用化学研究所，江西省南昌市 330029；2.江西农业大学，江西省南昌市 330045）摘要：以硅烷偶联剂KH560作为改性剂，采用化学合成方法合成了KH560改性酚醛树脂。
通过傅里叶变换红外光谱、热重分析以及力学性能测试研究了硅烷偶联剂KH560对酚醛树脂热性能和力学性能的影响。
结果表明：当w（KH560）为2.5%时，改性酚醛树脂在318 ℃时开始分解，树脂质量损失约为17.0%，耐热性能较好；与改性前相比，改性酚醛树脂的拉伸强度提高了32.9 MPa，冲击强度提高了4.03 kJ/m2，力学性能得到了改善。
KH 压力传感器技术数据说明书
KHPRESSURE TRANSMITTERU A RMain Features• Ranges: from 4 to 1000 bar• Nominal Output Signal:0...10Vdc (3 wires) / 4...20mA (2 wires)0.5...4.5 v ratiometric• Compact size• Wetted parts: Stainless steel• SIL 2 certified according to IEC/EN 62061:2005KH transmitters are based on film sensing element deposited on stainless steel diaphragm.Thanks to the latest state of the art SMD electronicsand compact all stanless steel construction, these pro-ducts are extremely robust and reliable, with SIL2 certi-fication supplied as standard.KH transmitters are suitable for all industrial applica-tions, specially on hydraulics (presses, pumps, powerpack, fluid power,etc.) with severe conditions usuallywith high level of shock, vibration, pressure and tem-perature peaks, as typical for mobile machines envi-ronment.FS = Full scale1)Incl. Non-Linearity, Hysteresis, Repeatability, Zero-offsetand Span-offset tolerance (acc. to IEC 62828-2)2)The operating pressure range is intended from 0.5 to100% FS3)Time within which the rated performance ia achievedPRESSURE RANGESRANGE(Bar)461016202540601001602002504006001000 Overpressure(Bar)812203240508012020032040050080012001200 Burst pressure(Bar)16244064801001602404006408001000150015001500 MECHANICAL DIMENSIONSELECTRICAL CONNECTION - ConnectorsELECTRICAL CONNECTION - Connection diagramsLOAD DIAGRAMPRESSURE PEAKS PROTECTIONSIL CERTIFICATION (Safety Integrity Level) – FUNCTIONAL SAFETYEXTENSION CABLESGEFRAN spa via Sebina, 7425050 PROVAGLIO D’ISEO (BS) - ITALIA tel. 0309888.1 - fax. 0309839063Internet: DTS_KH_08-2019_ENGGEFRAN spa reserves the right to make any kind of design or functional modification at any moment without prior notice.ORDERING INFORMATION。
HORIBA A-TEEM 分子指纹分析技术说明书
ELEMENTAL ANALYSISFLUORESCENCEOPTICAL COMPONENTSCUSTOM SOLUTIONSSPR IMAGINGAqualog®A-TEEM TMIntroducing the NEW HMMP tool for easybatch regression and discrimination analysis ofAqualog A-TEEM dataHORIBA’s patentedA-TEEM molecularfingerprinting isan ideal opticaltechnique forproductcharacterizationinvolvingcomponent quantification and identification. The HMMPAdd-In tool, powered by Eigenvector Inc. Solo, ideallycomplements the A-TEEM by supporting the developmentand batch wise application of methods for an unlimitednumber of component regression models as well asdiscrimination models. The HMMP breaks the time- andlabor-consuming barrier of analyzing individual modelsand collating results into a cohesive report to meet therequirements of industrial QA/QC applications. TheHMMP tool facilitates administrator level method modeldevelopment but more importantly push-button operator-level application and report generation.The HMMP tool is exclusive to the Aqualog A-TEEM andsupports enhanced model robustness by combining theabsorbance and fluorescence excitation-emission matrix(EEM) data using the Solo Multiblock Model tools! HMMPincorporates a direct, exclusive link to the Aqualog’s batchfile output directory for trouble-free file browsing andautomatic concatenation of absorbance and EEM data aswell as all model-dependent pre-processing.The HMMP tool mates seamlessly with data collectedusing the Fast-01 autosampler as well as any othersampling method that employs the Aqualog SampleQtoolbox.The HMMP tool supports an unlimited number ofregression models in a given method to providecomprehensive reports of all parameters of interest.Discrimination model methods with multiple class groupsare also supported to facilitate product characterizationas functions of unique compositions and component orcontaminant threshold concentrations among other QA/QC scenarios. The HMMP tool can employ a wide rangeof algorithms for discrimination and regression includingPrincipal Components Analysis (PCA), Partial LeastSquares (PLS), Artificial Neural Networks (ANN), SupportVector Machine (SVM) and Extreme Gradient Boost (XGB).Key applications supported include wine quality chemistry,water contamination and pharmaceutical productidentification and composition among many others.Key Features and Benefits• Easy, Rapid Operator Level Analysis• Facilitated Administration of Method Model Developmentand Editing• Complete Parameter Profile and Classification Reports• HMMP Add-In Fully Integrated into Eigenvector Inc.Solo/Solo+Mia and Exclusively Activated and Supportedby HORIBA Instruments Inc.• HMMP Reports include all required parameter informationand are saved in a comma separated format for LIMSsystem compatibility.• The HMMP tool is provided with ample online Helpsupport powered by the Eigenvector Inc. Wiki platformand HORIBA’s fully featured user manual.Aqualog A-TEEM Spectrometerwith FAST-01 AutosamplerPowered by Solo Predictor software fromEigenvector Research, IncorporatedHMMP SpecificationsTo learn more about theA-TEEM molecular fingerprinting technique, applications and uses of this autosampler, refer also to *******************/scientificUSA: +1 732 494 8660 France: +33 (0)1 69 74 72 00 Germany: +49 (0) 6251 8475 0UK: +44 (0)1604 542 500 Italy: +39 06 51 59 22 1 Japan: +81(75)313-8121 China: +86 (0)21 6289 6060 India: +91 80 41273637 Singapore: +65 (0)6 745 8300Taiwan: +886 3 5600606Brazil: +55 (0)11 2923 5400 Ot h er:+33 (0)1 69 74 72 00The HMMP user interface facilitates method development and selection, fully articulated data file browsing with data integrity warnings and push-button report generation.。
样车试制流程
查看完整版本: [-- 样车试制流程--]<< 1 2 Pages: ( 2 total )sicar07-09-12 10:53样车试制在整车的正向开发中，具有非常重要的意义。
我就个人理解把样车试制流程给大家交流下。
希望大家多多拍砖。
1、样车试制需要的条件数字样车的冻结。
车身钣金件的60%工装件的到件内外饰等快速成型件的备件动力总成、底盘等样件到件(国内好像没有开发底盘的，都是沿用)2、试制过程第一辆样车的周期最长。
大概三至四周的时间。
主要检验零部件的配合关系。
剩余的车辆时间就比较短了。
在此期间需要设计工程部门（车身、内饰、外饰等等）介入，现场跟踪解决问题。
同时还有质量管理、工艺人员等职能部门的介入3、试制结束样车制造的冻结。
解决在试制中出现的各种问题（配合问题，性能问题）。
然后进行小批量生产，然后是真正的投放市场，大批量生产。
请大家多多发表自己意见。
wltsha07-09-20 09:26 要是能用标准流程图的形式表达出来,那将会更完美.wltsha07-09-21 13:41流程,工程体系呀.例如同步工程,等最后都会落实到体现表格.图表等比较直观的事物上来,要不就太空泛了.我举几个例子,FEMA就是一张表格,上面体现出来了FEMA的精髓.,APQP,也是最后落实到各种表格,流程图上,.同步工程也是,落实到各种反馈表格上,这方面的例子很多.,希望大家在讨论流程,或者工程这些概念,能做些表格.流程图,就说明已经进入正轨了.wltsha07-09-28 15:23现在我们公司就在整理流程,用标准的流程框架.有设计规范,各个环节的管理用表格,等,这个是开发环节非常重要的内容,最好这些东西,才能使开发变的更容易.,wuyou81090207-10-13 13:45 看来各个公司都在开发新车,都进试作阶段了wenjuntang8107-11-04 22:14 以前不懂，长见识了。
yuecai121607-12-18 00:07 国内现在能做整车正向开发的好像还没怎么听说过。
KH-00目录
文件编号版次文 件 名 称备 注KH-0B0修订页
KH-00B0目录
KH-01B0接管验收工作细则
KH-02B0管理处入驻工作细则
KH-03B0客户入(退)伙管理程序
KH-04B0客户资料管理
KH-05B0装修管理程序
KH-06B0解答客户咨询技巧
KH-07B0客户投诉受理规程
KH-08B0客户回访作业指导书
KH-09B0租赁管理程序
KH-10B0钥匙管理规定
KH-11B0物业管理费用收取规程
KH-12B0车辆出入IC卡办理规程
KH-13B0住户智能卡办理规程
KH-14B0智能卡挂失/报损/补办规程
KH-15B0客户搬入搬出管理规程
KH-16B0停水停电管理作业指导书
KH-17B0商业网点﹑广告招牌管理规程
KH-18B0丽江豪园有偿服务项目收费标准
KH-19B0前台接待作业指导书
KH-19前台接待作业指导书KH-20B0社区文化活动管理规程KH-21B0客户维修作业指导书KH-22B0建筑物巡查管理要点KH-23B0装修管理规定。
全转录组测序分析精子发生中RNA结合蛋白质的动态表达
基础医学与临床Basic & Clinical MedicineJune 2021Vol.41 No.62021年6月 第41卷第6期文章编号：1001-6325 ( 2021) 06-0825-06研究论文全转录组测序分析精子发生中RNA 结合蛋白质的动态表达李 凯，邙新雨，邹定峰，李梦真，缪时英，王琳芳，宋 伟**收稿日期：2021-03-29 修回日期：2021・04-16基金项目：国家自然科学基金(31970794,32000586)* 通信作者(corresponding author ) : songwei@ (中国医学科学院基础医学研究所北京协和医学院基础学院生物化学与分子生物学系医学分子生物学国家重点实验室，北京100005)摘要：目的系统解析RNA 结合蛋白质(RBPs)在小鼠精子发生中的动态表达全貌、阶段特异性及协同表达模式，并预测其潜在调控作用。
方法整合6种类型生精细胞的全转录组测序数据，分析精子发生全程差异表达的RBPs ；利用时间序列分析软件(STEM)分析差异表达RBPs 动态表达模式；利用加权基因共表达网络分析(WGCNA)鉴定精子发生中协同表达的RBPs ；通过ClusterProfiler 工具分别对差异表达以及协同表达的RBPs 进行GO 功能富集分析。
结果精子发生中共鉴定519个阶段特异表达的RBPs,并具有7种动态表达模式，其中减数分裂时期的RBPs 占比最高；GO 分析显示RBPs 主要参与mRNA 选择性剪接、加工或翻译过程;WGCNA 分析获得246个共表达RBPs,其中减数分裂时期共表达RBPs 占比最高。
结论RBPs 在精子发生中呈现阶段特异性，并且以协 同表达模式发挥调控作用。
其在精子发生早期阶段参与RNA 加工或剪接等过程，而在后期阶段参与核糖体组装或RNA 翻译等过程。
关键词：RNA 结合蛋白质;转录组;共表达;精子发生中图分类号:Q28文献标志码:ADynamic expression of RNA-bindingproteins in spermatogenesis based on RNA-seqLI Kai, MANG Xin-yu, ZOU Ding-feng, LI Meng-zhen, MIAO Shi-ying, WANG Lin-fang, SONG Wei *(State Key Laboratory of Medical Molecular Biology , Department of Biochemistry and Molecular Biology, Institute ofBasic Medical Sciences Chinese Academy of Medical Sciences , School of Basic Medicine Peking Union Medical College ,Beijing 100005, China)Abstract : Objective To systematically characterize the dynamic expression pattern , stage specificity and co-ex pression pattern of RNA-binding protein ( RBPs) and to predict their potential regulatory role in mouse spermato genesis ・ Methods The whole transcriptome sequencing data of six spermatogenic cells types were integrated for an alyzing the differentially expressed RBPs during spermatogenesis ； STEM was used to analyze the dynamic expressionpattern of RBPs ； WGCNA was used to identify the RBPs co-expressed pattern ； The differentially expressed and co expressed RBPs were analyzed by the ClusterProfiler tool for GO function enrichment analysis. Results A total of 519 stage-specific RBPs were identified during spermatogenesis , and there were 7 dynamic expression patterns , ofwhich RBPs at the meiotic stage accounted for the highest proportion ； GO enrichment analysis showed that RBPs were826基础医学与临床Basic&Clinical Medicine2021.41(6) mainly involved in the selective splicing,processing or translation of mRNA；WGCNA analysis showed that246 RBPs were co-expressed,among which RBPs at the meiotic stage accounted for the highest proportion.Conclusions RBPs exhibit stage specificity and play a regulatory role in spermatogenesis with a coordinated expression mode.It functions mainly in the process of RNA processing or splicing in the early stage of spermatogenesis,and is have significant impact on the process of ribosome assembly or RNA translation in the later stage.Key words:RNA-binding proteins;transcriptome;co-expression；spermatogenesis精子发生是从精原细胞发育成为成熟精子的一个复杂有序的连续细胞分化过程，主要分为3个时期:精原细胞有丝分裂期、精母细胞减数分裂期和精子形成期⑴。
Quantabio repliQa
Quantabio, 100 Cummings Center Suite 407J, Beverly, MA 01915IFU-115.1 Rev01repliQa™ HiFi Assembly MixCat. No. 95190-010 95190-050Size:10 reactions 50 reactionsStore at -25°C to -15°CDescriptionThe repliQa™ HiFi Assembly Mix simplifies the construction of recombinant DNA through the simultaneous and seamless assembly of multiple DNA fragments possessing terminal regions of sequence overlap in a single, isothermal reaction. Similar in principle to the Gibson Assembly ® Method 1, the high efficiency repliQa HiFi Assembly Mix is ideal for a range of genetic engineering applications including routine molecular cloning, site-directed mutagenesis, assembly of large constructs for synthetic biology applications, and the construction of diverse sequence libraries for directed evolution studies. The concentrated, two-component format allows flexibility in design of assembly reactions and compatibility with less concentrated DNAsamples. The repliQa Mix has been optimized for use with a total input quantity of DNA fragments in the range of 0.03 to 0.5 pmols. The assembly of up to six DNA fragments is recommended, though the repliQa Mix has been successfully used for more complex assemblies.Double stranded DNA fragments for assembly can be generated by PCR amplification, chemical synthesis, or isolation of restriction fragments. When working with fragments PCR amplified from plasmid vectors, the included DpnI restriction endonuclease can be used for selectively digesting methylated, residual plasmid DNA to reduce background transformants. The repliQa mix is directly compatible with most common E. coli cloning hosts and generally provides a high yield of accurately assembled product.The DNA assembly occurs through the actions of three enzymes:• A non-thermostable 5' to 3' exonuclease that partially eliminates one strand of a DNA duplex to expose complementary overlap regions forhybridization.• A high-fidelity thermostable polymerase that fills the gaps remaining between the hybridized fragments of the overlapping regions.• A thermostable DNA ligase that covalently seals the resulting nicks at fragment junctions, generating double-stranded, assembled DNA moleculessuitable for transformation of cells.ComponentsReagent Description95190-01095190-050 repliQa HiFi Assembly Enzyme Mix Optimized formulation of enzymes for 5’-endresection, high fidelity 3’-end extension, and nick sealing.1 x 0.02 mL1 x 0.10 mLrepliQa 10X Assembly Reaction Buffer 10X reaction buffer containing dNTPs, magnesium, and cofactors.1 x 0.1 mL 1 x 0.50 mLDpnI (20 U/µl)Restriction endonuclease for the (optional) post-PCR digestion of residual unamplified plasmid template.1 x 0.05 ml 1 x 0.25 mlStorage and StabilityStore kit components in a constant temperature freezer at -25°C to -15°C upon receipt. For long term buffer storage (> 30 days) store buffer at -70°C. Refer to the product label or lot-specific Product Specification Sheet (PSF) available at /resources for applicable expiration date.A general diagram of assembly cloning is shown below:Additional reagents and materials that are not supplied• PCR-Grade, nuclease-free water (do not use DEPC-treated water)• High Fidelity DNA Polymerase (Enzymatics VeraSeq TM 2.0, P7511L or equivalent)• A heat block, thermocycler, or water bath capable of holding a temperature of 50 ± 2°C for one hour. • PCR or microcentrifuge reaction tubes.• PCR product purification kit (QIAGEN ® QIAquick ® PCR Purification Kit, 28104 or equivalent). •Competent E. coli cells and accessories as recommended by manufacturer.Before you begin• Design the DNA fragment sequences and assembly strategy. Guidelines are given in Appendix 1.• (Optional) Treat PCR reaction with DpnI if plasmid DNA was used as template for generating DNA fragments to be assembled. (Appendix 2).•(Recommended) After determining PCR fragment or restriction endonuclease-digested fragment size and purity by agarose gel electrophoresis, purify using a spin column-based cleanup or other method. This step is not required but is highly recommended to achieve highest efficiency of fragment assembly.• Measure the concentration of each isolated DNA fragment by absorbance at A 260 or by using a fluorometric quantitation reagent. Agarose gel electrophoresis with mass-calibrated size standards can also be used to quantify fragment mass and quality simultaneously. • Calculate the number of picomoles of each fragment using the following formula:pmols = (weight in ng) x 1000/(bp x 662).• Determine the number of pmols of each fragment to add to the assembly reaction. For cloning, highest efficiencies are achieved with 0.02 to 0.04 pmols of linear vector fragment (50 to 100 ng of 4 kb vector) and 2 to 8-fold molar excess of inserts. • Prepare outgrowth medium and culture plates with appropriate antibiotics for plasmid selection.•Equilibrate the heat block, thermal cycler, or water bath to 50°C for incubation of the assembly reactions .Protocol1. Thaw the repliQa HiFi Assembly Kit components, briefly vortex to mix, and place on ice.2. For each assembly, add reaction components in the order listed in the table below to chilled reaction tubes.The optimal amount of enzyme mix to add per assembly reaction depends on the total quantity of DNA fragments present.ComponentRxn. component volumes (µl) for varying amounts of total DNA≤ 0.125 pmol> 0.125 pmol but ≤ 0.25 pmol > 0.25 pmol Nuclease-free water(17.5 – X) µl (17.0 – X) µl (16.0 – X) µl repliQa 10X Assembly Reaction Buffer 2.0 µl 2.0 µl 2.0 µl DNA fragmentsX µl X µl X µl repliQa HiFi Assembly Enzyme Mix 0.5 µl 1.0 µl 2.0 µl Total volume20 µl20 µl20 µl3. Incubate reactions at 50°C in heat block, thermal cycler with heated lid (set to ~60-80°C), or covered water bath for 1 hr. Hold assembled product mix at 4°C until ready to proceed with transformations. If transformations cannot be performed on the same day, reactions can be stored at -20°C for up to one month.4. Competent E. coli should be transformed, recovered, and plated as per manufacturer guidelines or standard lab practices. Note: If electroporation is to be used for transforming cells, we recommend first diluting the assembly reaction 1:5 in high purity water. There is no need to dilute the assembly reactions prior to transformation of chemically competent cells.5. (Optional) Analyze a portion of the remaining assembly reaction by agarose gel electrophoresis. If DNA fragment assembly occurs properly, a ladder of higher molecular weight DNA bands would be generated.Note: For reactions using three or fewer fragments the incubation time in step 3 can be shortened to 15 minutes.Appendix 1 – Guidelines for Designing DNA Fragments for Assembly1.When designing the DNA fragment sequences and assembly strategy, allow for a region of sequence homology between adjacent DNA fragments.Be sure to avoid regions of repeated bases or repeated short DNA motifs in the design of these overlaps where possible. Regions of secondary structure such as hairpins or stem loops should also be avoided.2.The kit is optimized for the assembly of fragments with overlap regions between 15 – 60 bp. It is recommended that the overlaps be at least 20bp with a minimum of 25% GC content, however overlaps of 30 bp or longer size will provide higher efficiency assembly reactions.3.For generating PCR fragments to be assembled, design primers with a 5’ segment of homology to the adjacent fragment or vector. If the adjacentfragment is also generated by PCR amplification, the overlap can be split between two primers if desired. The 3’ segment of primers should contain sequence specific to the DNA target of interest. Amplify targets using a high-fidelity thermostable DNA polymerase such as VeraSeq 2.0 (Enzymatics, P7511L) or equivalent per manufacturer instructions.4.When designing synthetic gene fragments for assembly, ensure that the 5’ and 3’ segments contain regions of homologous overlap sequencebetween adjacent gene blocks, PCR fragments, or isolated restriction fragments.5.For site-directed mutagenesis applications, the assembly strategy should be designed such that the mutation of interest is centered betweenadjacent PCR fragments. Design the PCR primers as with the standard fragments above, except that the mutation (substitution, insertion, or deletion) should be included within the 5’ segments for both of the adjacent fragments.6.When designing DNA fragments to be assembled with isolated restriction fragments, be aware that any 5’ overlaps from staggered restriction cutswill be eliminated because of the 5’-->3’ nuclease present in the assembly mix, and so should not be included in the measurement of overlap size. If desired, design the 5’ overlap segment of the adjacent fragment to either preserve or eliminate the restriction site.Appendix 2 – DpnI treatment to remove residual plasmid DNAWhen plasmid vector is used as PCR template to generate a fragment for assembly, it is recommended that the reaction be treated with DpnI to eliminate residual methylated plasmid prior to setting up the assembly reaction.1.Add 1 µl DpnI (20U) directly to the PCR reaction (50 µl) following amplification of fragment.2.Incubate at 37°C for 1 hr.3.Heat inactivate DpnI by incubation at 80°C for 20 min.4.(Recommended) Purify the fragment using a spin column-based PCR purification kit.Quality ControlThe repliQa HiFi Assembly Mix is functionally tested for assembly of three 1-kb PCR fragments into 2kb and 3 kb products.The individual components of the repliQa HiFi Assembly Mix are tested to be free of contaminating DNase and RNase.Limited Label LicensesThis product was developed, manufactured, and sold for in vitro use only. The product is not suitable for administration to humans or animals. SDS sheets relevant to this product are available upon request.References1. Gibson, D.G., et al. (2009). Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat. Methods 6, 343-5.。
ProjectDescripti...
Sector Agro and Food ProcessingSub- sector Agri-infrastructureProject No AF- 01Project/Product Pack-house Project for BananaProject DescriptionBanana pack house project will provide all the technical facilities for pre and post harvest procedures, so as to maintain consistent quality for export of the product.These facilities can also be used for supply of high quality Bananas in the domestic market and to Banana processing units in Gujarat.The proposed unit will have tie-up for Banana procurement from the farms, and will facilitate Banana sorting, grading,cleaning line, Banana pre-cooling, Controlled atmosphere Banana cold storage, Banana ripening chambers and refer transportation system for Banana supply to export market or to the exporting sea ports till the material is loaded in refer sea containers for exports.Mechanized Banana handling system using wire rope conveyors for hand collection of green banana from the farm can also be included as project component,where large quantity of green bananas have to be hand picked from an area deprived of direct access to plantation or requires movement through high density plantation.Project ConceptThe proposed project is an important link in the complete Supply Chain Management of Banana marketing in export and domestic markets,while providing necessary support to Banana growers of particular area for the development of Banana markets. The project will provide required impetus in the form of physical infrastructure and technological input to the Banana growers and exporters,which in turn will develop good quality banana that can be exported from the region.Market & Growth Drivers∙Global production of Banana in the year 2004 was estimated at around 70 million tons. India is one among the10 major banana producing countries which together accounted for about 75% of total World banana production in the year 2004.Production as well as trading of Bananas is highly concentrated in a few Asian and Latin American countries. This concentration of banana production has intensified over a period of time and is showing a different regional distribution.∙The Latin American and Caribbean region dominated Banana Production up to eighties, while the Asian region took the lead in Banana production during the nineties. African production levels have remained relatively stable during that period. Distribution of World Banana production as per FAO statistics, between 2001 to2005is graphically shown in the following figure.S ource: Food and Agriculture Organization of the United States (FAO)∙India contributed23%to the global banana production and 11%to the total area under cultivation.Rising Banana productivity in Gujarat as well as India is becoming a matter of concern for post harvest facility of storage and handling.∙The major countries to which India exports banana are U.K., Saudi Arabia, Kuwait, UAE, Oman, Qatar, and Yemen Arab Republic.∙Technological change by the developed countries in the post harvest storage methodology boosted global banana trade which registered rapid growth in the last few years and has reached INR 436 billion.Banana Trade StatisticsSr.Years Quantity (MT)Value (INR Lacs) No12003-20041426.41166.2122004-2005924.4161.6532005-20062033.8199.2342006-2007(Apr- Jun)1740.31200.48Source: Department of commerce, Government of IndiaGrowth Drivers∙India does not export even1% of the total banana production and thus, there is a vast potential for increasing this quantity, provided a Banana pack house system comes up.∙Indian Banana production has increased substantially in the last few years, due to increased adoption of tissue culture plants which gives higher yield, consistent quality and production for longer period of year.∙There is a huge potential for export to EU countries and CIS countries. India already exports Bananas to Germany, France, U.K., but still needs to go a long way to meet strict quality standards.Why Gujarat?∙In India,Gujarat is one of the leading states in banana production, with the highest productivity of 40 MT/ ha. The State production in the year 2004-05 was1.98 million tons and is expected to grow with availability of canal irrigation & adoption of tissue culture plants as planting material.∙Government of Gujarat,being aware of the current situation of Banana export, offers 6% back end interest subsidy with a ceiling of INR4000million for Banana Pack house project.∙Gujarat has availability of technically trained man power for pack house operation at economical cost.∙Gujarat has well developed transport infrastructure facilities like rail & road network and availability of world class ports like Kandla, Mundra and Pipavav having required container cargo handling facilities.Raw MaterialsThe Area and production of Bananas in Gujarat for the last 5 years period are summarized in the following Table:Area and Production of Bananas in GujaratSr. No.Year Area (’00 Hectares)Production (MT)12001-0233139115433022002-0335187140307732003-0442909176090142004-0546347197925752005-06492342498776Source: Department of Horticulture Statistics, Gandhinagar, Government of GujaratAs observed from the above table Banana production in Gujarat has seen an escalating trend, in terms of area and production in the last five years. This production increase can be attributed to enhancement in average yield per hectare.The major Banana producing districts considering area under cultivation and production are Anand, Surat, Baroda, Bharuch, Narmada and Kheda.Technology/ProcessBeing a core infrastructure project it will require Banana procurement system from farms, then cleaning, sorting, grading,packing line and pre-cooling them under controlled atmosphere cold storage,with Banana ripening and Logistic system to carry them to ports.In case the operational area of the proposed pack house is having more than 40 km periphery, it is suggested to have mobile pre-coolers so as to eliminate any damage to the crop from farm heat before being brought to the pack house.Mechanized Banana handling system, as used in LatinAmerican countries, can also be planned, where Banana hands have to be picked from large farm areas or from Banana farms which cannot be approached due to high density.In Gujarat, the horticulture department as well as the state agriculture universities would provide necessary extension services and technological inputs to the farmers or their co-operatives, for getting quality Banana crop from the region.Suggested Project Capacity and Project CostThe proposed Banana pack-house would have capacity of handling and storing 1000 MT of Banana at any point of time. It would be equipped with a ripening chamber to cater to the need of the domestic market. The proposed pack-house would store Bananas for the duration of 2-3 months.The estimated project cost would be INR 45 million (US $ 1 million).Estimated Project cost & Means of financeSr.Cost of project INR in millionNo.1Land and Land development 3.002Building10.003Plant & Machinery (1000 MT Storage)19.004Misc. Fixed Assets 2.505Preliminary & preoperative incl. technology 2.506Provision for contingencies 2.00 Fixed Capital cost39.007Margin Money for working capital 6.00 Estimated Block Capital Cost of Project45.00Means of Finance8Promoters contribution (Debt equity ratio is 2.5:1)15.009Term loan30.00 Total Means of Finance45.00As indicated above, the proposed project will require an approx6000 sq. mt of land with an proposed built up area of2000 sq. mt.The Banana pack house unit is proposed to have storage capacity of 1000 MT and considering 3 storage cycles in a year its effective storage capacity will be 3000 MT per annum. The total fixed cost of the project is estimated at INR 39 million and INR6 million is the working capital margin, which will make total block capital cost to INR 45 million. The unit being proposed to cater to domestic as well as International demand, is suggested to have a Debt equity ratio of 2:1. Thus, the estimated term loan amounts to INR 30 million and Equity at INR 15 million. Suggested LocationBanana pack-house could be set up at various places in Central and South Gujarat Regions in the district of Kheda, Anand, Part of Panchmahal,Vadodara, Surat, Narmada, Bharuch,and Valsad. Plant and MachineryThe list of main plant and machineries for the proposed Banana pack house is summarized in the following table:List of Plant and Machinery Sr.No.Particulars Quantity Supplier1 Banana Brush washer2Global Agri Tech Engineer, Vadodara Process Masters, MIDC Pimpri, Pune-Maharashtra2Banana Inspection Conveyors1Global Agri Tech Engineer, Vadodara3Pre coolers and Pre coolingchambers 25 MT / day4Frick India Ltd, New Delhi & Mumbai4Banana Cold Storage 250 MT each4Frick India Ltd, New Delhi & Mumbai5Banana Ripening chambers 10 MTeach3“Lock Sock” Ravin Impex (P) Ltd,Ahmedabad-3800096Heavy duty Plastic crates forBanana storage and handling20000Neelkamal Plastics Mumbai7Ammonia Compressors for Precooling and Cold Storage4Frick India Ltd, New Delhi & Mumbai8Atmospheric condensers2Frick India Ltd, New Delhi & Mumbai9HT / LT Electrical system with DGset for stand by powerLot Kirloskar Electricals Ltd-Ahmedabad10Pallet handling truck1Godrej & Boyce Ltd, Mumbai11Refer insulated vans for transport ofBanana from farms & to market2Tata Motors /Ashok Leyland & CarrierAircon Ltd-DelhiUtilitiesThe proposed Banana pack house will have electrical connected load of 120 HP and utilization will be approx.80 HP on a regular basis. The unit will require approx.20 KL water for Banana cleaning and cooling system of refrigeration unit proposed for pre-cooling and cold storage unit.Man power requiredThe proposed project will have total manpower requirement of 20 persons. This will include 3 managerial posts, 3 supervisory posts, 6 operators, 3 engineering maintenance staff like electrician, mechanical foremen and 5 accounts,administrative and security staff.Project Time LineThe proposed project will have cumulative implementation period of 10- 12months of which 5 to 6 months would entail obtaining the required clearances from various authoritiesFinancial IndicatorsBased on the profitability projections worked out for the proposed project, key financial indicators are as summarized below:Key Financial IndicatorsSr. No.Particulars1st Year2nd Year3rd YearA Break-Even Point53.4%41.0%30.1%B Debt-service Coverage Ratio 1.50 1.96 2.83C Average DSCR 2.10D Return on Investment (ROI)26.9%46.4%68.1%E IRR26%As perceived from the Project cost and Means of finance table, the suggested Debt Equity Ratio for the proposed project is2:1. The IRR (Internal Rate of Return) for the proposed project is approx. 26% projected for a period of 10 years.Clearances RequiredFilling of Industrial Entrepreneur’s Memorandum (IEM) with the Secretariat of Industrial Approvals (SIA), Department of Industrial Development, Ministry of Industry, New Delhi.Registration with Ministry of Food Processing Industries (MOFPI), through state nodal agency GAIC, to avail benefits of scheme for food processing industry- under Agri Infrastructure Project category Approach state office of National Horticulture Board for availing incentives under the scheme for Banana Pack house and other post harvest infrastructure facilities including cold chain transportation Critical AspectsIt is very much important to examine critical aspects of such project at the planning stage to overcome main hurdles of the project and to undertake successful implementation. These are as under:The Success of such export oriented pack house is contingent on consistent supply of good export quality materials in required quantity, as per international standards.To achieve smooth exports, clearly defining the pre-harvest and post harvest procedures, Quality norms for different markets, training of growers, filed staff and pack house staff regarding quality standards are very vital.Efficient operation of post harvest facilities at pack house as well as transit handling system for economical operation is important.Development of good vendors for supply of packing cartons, Ethylene gas,regular service of post harvest equipments and reefer transportation vehicles, and stand by utility supply equipments.Agencies to be Contacted Industrial Extension BureauGujarat Agro Industries Corporation Ltd. Mott MacDonald India。
分层采油用丢手压力计算方法
分层采油是解决牙刷状油藏、叠层状油藏、高含水油藏等层间干扰[1-3]，提高最终采收率的有效手段之一，如分采层段数不受限制的压力波控制分层配产、过环空缆控分层采油、无线对接式缆控分层采油、振动波控制分层采油等技术[4-6]。
实际上，在工业4.0时代，智能油藏、智能决策、智能找堵水等基础上的分层采油井下丢手结构管柱也不可或缺[7]。
投球丢手则是分层采油过程中必要的技术手分层采油用丢手压力计算方法*付亚荣陈劲松郭志强刘万斗马永伟孙佰球胡俊平王晓军刘志刚钱洪霞王嫱（中国石油华北油田分公司）摘要：丢手工具是油气开采领域中的重要器具，具有实现封隔器、尾管等井下工具或管柱的送入及分离功能，在分层采油中起关键作用。
为解决分层采油丢手压力设计凭经验进行的弊端，达到既保证投球丢手的需要，又不造成管柱蠕动的目的，发明了一种分层采油用丢手压力的计算方法。
基于Hertz 接触理论，首先计算丢手上方水柱压力，然后分别对钢球所受的法向力、钢球与球座的接触长度、钢球与球座之间的最大接触压力进行计算，最后得到丢手压力。
经现场50余口油井应用表明，井下施工一次成功率达100％，助力30余口油井分层开采，单井日产油量平均增加3.4t，单井日产水量平均降低5.9m 3，百米吨液耗电平均降低0.107kWh。
关键词：分层采油；丢手；压力；Hertz 接触理论DOI :10.3969/j.issn.2095-1493.2022.07.020Calculation method of release pressure for separate-zone productionFU Yarong，CHEN Jinsong，GUO Zhiqiang，LIU Wandou，MA Yongwei，SUN Baiqiu，HU Junping，WANG Xiaojun，LIU Zhigang，QIAN Hongxia，WANG Qiang Huabei Oilfield Branch Company of CNPCAbstract：Release tool is an important tool in the field of oil and gas production.It has the function of feeding and separating downhole tools or pipe strings such as packer and liner.It plays a very important role in separate-zone production．In order to solve the disadvantage of experience in the design of re-lease pressure，meet the needs of ball throwing and release without causing string creep，calculation method of release pressure is invented．First，we should calculate the water column pressure above the release based on Hertz contact theory，then we should calculate the normal force on the steel ball，the contact length and the maximum contact pressure between the steel ball and the ball socket and finally we obtain the release pressure.More than 50oil wells have been applied on site and the first-time suc-cess rate of construction is 100%．It assists the implementation of layered production of more than 30oil wells and the average oil production of a single well is increased by 3.4tons while the average water production of a single well is decreased by 5.9m 3and the average power consumption is decreased by 0.107kWh．Keywords：separate-zone production；release tool；pressure；Hertz contact theory 第一作者简介：付亚荣，高级工程师，1987年毕业于重庆石油学校（油田应用化学专业），从事油气田开发技术研究与应用工作，************，**********************.cn，河北省辛集市华北油田第五采油厂，052360。
句子成分及基本句型
4. My brother and I go to school at half past seven in the morning and come back home at five in the afternoon. （并列主语和并列谓语）
一套作业本听写本作业本作文本一本纠错本一本纠错本一本笔记本至少够一学期用一本字典一本新概念英语第二册每周背诵一篇周背诵一篇一套作业本听写本作业本作文本一本笔记本至少够一学期用一本字典一本新概念英语第二册每?今天我背单词了吗
Evaluation only. ted with Aspose.Slides for .NET 3.5 Client Profile 5.2
Copyright 2004-2011 Aspose Pty Ltd.
Advice onEvlealauartnioinnognlyE. nglish
ted with Aspose.Slides for .NET 3.5 Client Profile 5.2 Copyright 2004-2011 Aspose Pty Ltd.
英语句C子o的py基ri本gh结t构2可00以4归-2纳0成11五种As基p本o句se型P及t其y 扩Lt大d.、组合、
省略或倒装。掌握这五种基本句型，是掌握各种英语句子结构的基 础。
• １．主语：主语是在句子中说明全句中心主题的 部分．一般由名词，代词，不定式，动名词或从 句充当．它的位置一般在句首． Evaluation only.
2013设备总台帐 machine list
36
051A04003 空 压 机 Air compressor
GA75+FF
设 备 台 帐 Machine List
主要性能 Main performance
切割厚度cutting thickness:40mm 工作台work table:4200*12000mm 电源power supply: M3*360MGC 操作系统operating system: VISION 51 切割厚度cutting thickness:35mmm 工作台work table:4000*12000mm 电源power supply: HPR260 操作系统operating system: Global
30
058A01006 喷漆线底漆通风设备 Primer areation
AL-KO
31
058A01017 喷漆线底漆通风设备 Primer areation
AL-KO
32
058A01018 小件喷漆通风设备 Primer areation
AL-KO
31
058A05001 滚筒抛丸机Shot blasting machine
2008.10 2006.04
9kW 11.5kW 3.4kW
6kW
昆山梅塞尔公司 Messer(Kunshan) 上海新力机器厂 Shanghai Xinli 靖江锻压机床厂 Jing Jiang
德国 Kasto-Germany 德国 HANS KALTENRACH-Germany 上海新力机器厂 Shanghai Xinli 天津埃佛茵机械有限公司 Tianjin AFM 徐州锻压设备厂 Xuzhou 锡山市堰桥机床厂 Xishan Yanqiao 泰安华鲁锻压机床有限公司 Tai An Hualu 出厂日期：2011.3 出厂编号：116052 瑞典 Round-Schwenden 意大利FACCIN Italy 出厂日期：2012.5.16 出厂编号：12044211-4186 安装日期：2012.7.9 荷兰 Gietart-Dutch 北京第一机床厂 Beijing No. 1 中捷摇臂钻床厂 Zhong Jie 中捷摇臂钻床厂 Zhong Jie 中捷摇臂钻床厂 Zhong Jie
光纤光学课后习题答案
5.3 Solution:
I don’t think so. MFD is a parameter using only for SMF which represents characteristic of a field distribution, while in MMF we use core diameter instead.
kh da w. co m
2 n12 - n2 and Δ =
5.2 Solution:
n1 - n2 , just let (NA) be 0.125, (n1 + n2 ) / 2
课
后
2 η = exp(- 2r 2 / w0 ) = exp(- 0.5) = 0.607
答
案 网
ì n(r ) = n1 ï ï í ï ï î n(r ) = n2
8.2 Solution
(a) a=62.5
μm ,x=3 μm ,
ww
(b) For
8.3 Solution n=1.467, λ=1310nm,w0=9.3um,x=0.5um, z=0.5um, φ=30’
(a) Loss lat = −10 lg[exp(−( x / w0 ) )] = 0.01dB
6.17 Solution:
ww
a. FWM is one of the major nonlinear phenomenon which can be described as follows: three waves copropagating over the same fiber generate the fourth wave, whose frequency is the combination of the three original wave frequencies. b. Because FWM has become the most sever limitation of a system’s transmission capacity.
氧化石墨烯聚甲基丙烯酸丁酯复合材料的热稳定性(论文)
Vol.35高等学校化学学报No.112014年11月 CHEMICAL JOURNAL OF CHINESE UNIVERSITIES 2466~2471 doi:10.7503/cjcu20140120氧化石墨烯/聚甲基丙烯酸丁酯复合材料的热稳定性张丹凤1,2,范楼珍3,郭瑞华3,樊择坛3(1.辽宁大学化学院,沈阳110036;2.黑龙江八一农垦大学理学院,大庆163319;3.北京师范大学化学学院,北京100875)摘要 采用3-氨基丙基三乙氧基硅烷偶联剂(KH550)对氧化石墨烯表面进行修饰,得到功能化氧化石墨烯(FGO),然后在引发剂偶氮二异丁腈(AIBN)作用下,以甲基丙烯酸丁酯(BMA)为单体进行原位聚合反应,得到功能化氧化石墨烯/聚甲基丙烯酸丁酯(FGO /PBMA)复合材料.通过红外光谱(FTIR)㊁X 射线光电子能谱(XPS)㊁扫描电子显微镜(SEM)和热重分析(TGA)等手段对复合材料进行表征.结果表明,所制备的FGO /PBMA 复合材料与聚甲基丙烯酸丁酯材料相比热稳定性显著提高,TGA 结果显示,FGO /PBMA 的最大失重区由PBMA 的267ħ升高到339ħ,同时对反应机理进行了探讨.关键词 氧化石墨烯/聚甲基丙烯酸丁酯;原位聚合;热稳定性;反应机理中图分类号 O645 文献标志码 A 收稿日期:2014-02-20.网络出版日期:2014-10-21.基金项目:国家自然科学基金(批准号:21073018,21233003)资助.联系人简介:张丹凤,女,讲师,主要从事纳米功能材料的修饰研究.E-mail:smile02df@范楼珍,女,博士,教授,博士生导师,主要从事电化学和纳米材料的制备及在燃料电池和传感器领域的应用研究.E-mail:lzfan@聚甲基丙烯酸丁酯(PBMA)具有高度的透明性㊁优良的光学性能㊁较好的力学强度和绝缘性能,耐水㊁耐油㊁耐碱等,特别是具有优异的相对伸长率和良好的黏结性能,适用于航空㊁航海设备的抗冲击透明件的胶合,还可以与其它高分子材料共混用于印刷品封面及用作涂料改性剂等[1].但其耐热性能差,只能在玻璃化温度下使用,且极易燃烧,大大限制了其应用.目前PBMA 热稳定性改善主要是采用阻燃剂或与其它有机物形成共聚物[2],而利用石墨烯(GP)的优异性能并将其添加到PBMA 中以改善其性能方面的研究鲜有报道.石墨烯是一种疏水性材料,在大部分溶剂中的相容性都较差;而氧化石墨烯(GO)表面存在大量的含氧基团,如羟基㊁羧基㊁环氧基等,可以通过共价或非共价的方法在石墨烯表面进行功能化修饰,得到石墨烯复合材料[3~5].Wang 等[6]在甲基吡咯烷酮(NMP)中剥离石墨得到石墨烯后原位引发聚合反应,制备GP /PMMA(PMMA =聚甲基丙烯酸甲酯)复合材料,提高了本体PMMA 的机械性能及稳定性.Pramoda 等[7]在氧化石墨表面修饰上十八胺(ODA)后再与甲基丙烯酰氯反应引入可聚合的碳碳双键,然后原位聚合甲基丙烯酸甲酯,得到共价修饰的聚合物材料.Pham 等[8]利用阳离子自由基引发剂自组装得到通过静电作用结合的聚甲基丙烯酸甲酯-还原氧化石墨烯(PMMA-RGO)复合材料.此外,还可以通过其它原位聚合反应制备与石墨烯复合的各种材料[9,10].Vallés 等[11]采用机械熔融共混的方法将氧化石墨烯引入PMMA 中,从而加固了PMMA.本文首先采用3-氨基丙基三乙氧基硅烷偶联剂(KH550)对氧化石墨烯进行表面功能化修饰,然后在引发剂作用下原位聚合甲基丙烯酸丁酯,使功能化的氧化石墨烯有效分散于聚合物本体中.对聚合物本体PBMA 热稳定性的考察结果表明,所得到的FGO /PBMA 复合材料的热稳定性相比于纯PBMA 有显著提高,在黏结剂㊁密封胶㊁有机玻璃㊁光学领域和生物医学工程等方面具有广泛的应用前景.1 实验部分1.1 试剂与仪器石墨烯(粒径1~5nm),南京先丰纳米材料科技有限公司;3-氨基丙基三乙氧基硅烷偶联剂(KH550),荆州市江汉精细化工有限公司;偶氮二异丁腈(AIBN),武汉盛世精细化学品有限公司;甲基丙烯酸丁酯(BMA),分析纯,天津市光复精细化工研究所;无水乙醇㊁N ,N -二甲基甲酰胺㊁浓硫酸(质量分数98%)㊁高锰酸钾和过氧化氢(质量分数30%)均为分析纯,国药集团化学试剂有限公司.SK1200H 超声清洗仪,上海昆山仪器有限公司;TG16-2台式高速离心机,湖南湘仪实验室仪器开发有限公司;IRAffinity-1傅里叶变换红外光谱仪(FTIR),日本岛津公司;JEOL JSM 6700F 场发射扫描电子显微镜(SEM),日本JEOL 公司;Multi Mode V SPM 原子力显微镜(AFM),美国VEECO 公司;ESCALab 250Xi X 射线光电子能谱仪(XPS),Thermo Fisher Scientific 公司;TGA /DSC1STARe 热重-差热分析仪(TGA-DSC),美国Mettler Toledo 公司.1.2 功能化氧化石墨烯(FGO )的制备采用Hummers 方法[12,13]以石墨烯(1~5nm)为原料合成氧化石墨烯.将30mg 氧化石墨烯置于15mL 无水乙醇中,超声分散30min,在装有冷凝管的三口瓶中磁力搅拌,升温至回流,滴入已溶解的5mL 偶联剂[V (偶联剂):V (乙醇)=1:5],继续搅拌并保持回流温度反应6h,取出部分反应液(5mL),离心并水洗3次,于60ħ烘干24h,制得FGO.1.3 原位聚合反应制备FGO /PBMA 复合材料在装有15mL FGO 反应液的三口瓶中通入N 2气约30min,搅拌,升温至回流温度,加入6mg 引发剂AIBN 并滴加15mL BMA 单体,保持回流温度继续反应6h,稍冷却后过滤,洗涤,于60ħ烘干,得到浅灰黑色固体,即为FGO /PBMA.2 结果与讨论2.1 红外光谱石墨烯在酸和高温处理下得到GO,然后再与聚合物本体在引发剂作用下进行原位聚合,相应特Fig.1 FTIR spectra of GO (a ),FGO (b )and FGO /PBMA (c )征峰的出现或移动可以从FTIR 谱图中发现.图1为GO,FGO,FGO /PBMA 的红外光谱图.从图1谱线a 中可以看出,氧化石墨烯中含有大量的亲水基团,如 OH, COOH, C O 等.其中,3300~3700cm -1处的宽吸收峰为氧化石墨烯中的 OH 及其吸收水分子中的羟基峰,1633cm -1处为芳基羧酸中 COOH 的特征吸收峰,而在1730cm -1左右出现的弱吸收峰为芳基羧酸中的 CO 伸缩振动吸收峰[14].图1谱线b 为GO 经过偶联剂KH550功能化后的红外谱图,其中3400~3200cm -1处为 NH 的特征峰,1660cm -1处为 NH 2的伸缩振动峰,羰基伸缩振动吸收峰由GO 的1730cm -1处移至1700cm -1处,说明偶联剂KH550中的 NH 与GO 中的 COOH 形成了酰胺键,偶联剂已与GO 发生作用.图1谱线c 为功能化后进行接枝反应所得产物的红外谱图,谱图中清楚地显示出了相关的特征峰.另外,在图1谱线c 中1096cm -1处为Si O Si 键的反对称伸缩振动峰,940,787,698cm -1对应于Si OH 键的伸缩振动,均为偶联剂KH550的特征峰.说明GO 与偶联剂KH550发生了缩合反应,KH550的功能化有机链与GO 通过酰胺键连接到一起.在1300~1400cm -1处出现的吸收峰为 CH 3和 CH 2 的C H 键的弯曲振动峰,2800~2900cm -1处出现的吸收峰为 CH 3和 CH 2 的C H 键的伸缩振动峰,说明功能化的GO 表面亦存在甲基丙烯酸丁酯.7642 No.11 张丹凤等:氧化石墨烯/聚甲基丙烯酸丁酯复合材料的热稳定性2.2 X 射线光电子能谱XPS 是研究石墨烯表面官能团的有力表征手段[14,15].图2为GO 和FGO /PBMA 的XPS 全谱扫描图.图2(A)中C 和O 元素的特征峰来自GO 表面存在的 COOH 和 OH 官能团,图中并没有明显的Si 和N 峰.图2(B)中则出现了较明显的Si 峰和N 峰,可以推测功能化的GO 与单体BMA 在引发剂AIBN 的作用下发生反应并成键,这与前面FTIR 的分析结果一致.Fig.2 Wide scan XPS of GO (A )and FGO /PBMA (B )图3为GO 的C 1s 谱(A),FGO /PBMA 的C 1s 谱(B)和N 1s 谱(C).图3(A)中284.70eV 处为C C 的信号峰,而285.06eV 处为C C 的信号峰,其中CC 属于GO 平面苯环的骨架和GO 边缘的修饰基团,C C 属于GO 平面内部缺陷和GO 边缘的修饰基团.另外,287.23,287.95,289.01eV 处分别对应C O,C O, COO 的结合能,说明氧化石墨烯的平面内和周围有大量的含氧基团存在.Fig.3 Narrow scan XPS of C 1s for GO (A )and FGO /PBMA (B )and N 1s for FGO /PBMA (C )从图3(B)中可以看出,在FGO /PBMA 的C 1s 谱中出现了C N 峰(285.97eV),C O,C C, COO 的结合能分别为286.53,284.59,288.78eV.表1中给出了相应峰的峰值及其相对比例.相比于图3(A)中GO 的相应峰值,FGO /PBMA 的峰值均有移动,且其中的C C,C C,C O 的峰高均有明显减小,说明GO 表面所含的含氧基团减少,这可能是由于GO 表面的 COOH 与KH550所含的 NH 2作用所致.图3(C)为FGO /PBMA 的N 1s 谱,显示有明显的 NHCO 峰,说明GO 表面存在C N 键,即在GO 表面引入了有机链.Table 1 Peaks and fractions of different C-containing groups in Fig.3(B )Bond E b /eV Fractions(%)Bond E b /eV Fractions(%)C C 284.5912.39C O 288.02 2.44C N 285.9728.51 COO288.78 2.08C C 284.9438.59C(epoxy)286.5315.992.3 结构表征由GO 的SEM 照片[图4(A)]可以看出,GO 呈均匀的片层状,由于GO 表面含氧官能团的存在使得表面出现褶皱堆积.图4(E)为GO 的AFM 照片.可以看出,GO 的片层厚度约为0.85nm,接近单层石墨烯的厚度.由图4(B)PBMA 的SEM 照片可以看出,聚合物PBMA 呈黏结状态,这可能是由于分子间发生聚合作用,单分子容易聚合成有机高分子长链引起的.图4(C)是FGO 的SEM 照片,由于偶联剂KH550的修饰作用使功能化的氧化石墨烯片层增厚,表面变得更光滑.图4(D)则是最终合成8642高等学校化学学报 Vol.35FGO /PBMA 复合材料的SEM 照片,由于聚合物与功能化的氧化石墨烯纳米片之间的π-π相互作用,使两者之间更容易发生反应,复合材料表面的褶皱明显增多,断面处有较明显的层状结构,但表面仍较光滑.图中显示复合材料分布均匀有条理,并不堆积,说明FGO 已经较好地分散到聚合物PBMA 本体中.SEM 结果与XPS 及FTIR 的分析结果一致.Fig.4 SEM images of GO (A ),PBMA (B ),FGO (C ),FGO /PBMA (D )and AFM image ofGO (E )with height profile (F )2.4 热重分析图5为GO,FGO,PBMA 和FGO /PBMA 的热重曲线.功能化石墨烯在聚合物PBMA 本体中分散效Fig.5 TGA curves of GO (a ),FGO (b ),PBMA (c )and FGO /PBMA (d )in N 2果的好坏直接影响复合材料的性能,而且PBMA 本体与FGO 界面相互作用的强弱也会受到影响[8].通过TGA 分析比较FGO /PBMA 复合材料与GO 和PBMA 的热稳定性能.实验在N 2气氛中进行,由室温开始升温至600ħ,升温速度为10ħ/min.从图5中可以看出,GO 是热不稳定的,其主要的失重区在160ħ左右,这主要是由于GO 表面存在大量的含氧基团如 OH, COOH 等所致,这些含氧基团在受热后以CO,CO 2,H 2O 等形式使GO 失重,而FGO 由于偶联剂KH-550与GO 表面的 COOH 作用,连上了有机链,相比于GO 热稳定性有所增加.聚合物PBMA 在200ħ开始失重,最大的失重区在267ħ左右,失重率为6.56%.而FGO /PBMA 复合材料相比于PBMA 热稳定性明显增强,于230ħ左右开始失重,最大失重区发生在339ħ左右,失重率为2.07%.由于FGO 片层均匀分散于聚合物PBMA 本体中,而PBMA 与FGO 之间有较强的界面作用力,因而所制备的FGO /PBMA 复合材料的热稳定性较PBMA 明显提高.2.5 反应机理基于前述研究结果推测了氧化石墨烯与有机物单体作用的反应机理.氧化石墨烯表面含有大量的含氧基团,如 OH 和 COOH 等,实验中硅烷偶联剂KH550中所含的氨基与氧化石墨烯表面的羧基进行酰胺化,在氧化石墨烯表面连上了硅烷长链,使氧化石墨烯功能化(Scheme 1).功能化的氧化石墨烯与单体BMA 在引发剂AIBN 的作用下发生原位聚合作用,氧化石墨烯经过功能化后表面接有硅烷长链,使其在有机相中的相容性增加,可以均匀分散于有机单体中,并与之反应成键(Scheme 2).9642 No.11 张丹凤等:氧化石墨烯/聚甲基丙烯酸丁酯复合材料的热稳定性Scheme 1 Functionalization ofFGOScheme 2 Reaction between BMA and FGO to produce FGO /PBMA3 结 论首先通过硅烷偶联剂KH550对氧化石墨烯表面进行功能化修饰,使修饰后的氧化石墨烯在有机相和无机相间的相容性明显增强,然后再进行原位聚合反应,在引发剂AIBN 的作用下,使单体BMA 与FGO 在一定条件下反应,所得FGO /PBMA 复合材料的热稳定性明显增强.PBMA 在200ħ开始失重,最大失重区发生在267ħ,而FGO /PBMA 复合材料从230ħ开始失重,最大失重区发生在339ħ.实验采用功能修饰的氧化石墨烯作为填充剂来改善PBMA 的热稳定性,反应可以一步完成,方法简单易行.参 考 文 献[1] Xiao W.D.,Cheng S.Y.,Adhesives and Sealants ,Chemical Industry Press,Beijing,2002,130 135(肖卫东,程时远.密封胶粘剂,北京:化学工业出版社,2002,130 135)[2] Wang X.L.,Miao C.Q.,Ju C.X.,Bao J.C.,Plas.Sci.Tech .,2009,37(1),56 59(王新龙,苗彩琴,鞠昌迅,包建春.塑料科技,2009,37(1),56 59)[3] Huang X.,Yin Z.Y.,Wu S.X.,Small ,2011,14(7),1876 1902[4] Chen Z.X.,Lu H.B.,Chem.J.Chinese Universities ,2013,34(9),2020 2033(陈仲欣,卢红斌.高等学校化学学报,2013,34(9),2020 2033)[5] Yu L.,Zhang Y.T.,Liu J.D.,Chem.J.Chinese Universities ,2014,35(5),1100 1105(余亮,张亚涛,刘金盾.高等学校化学学报,2014,35(5),1100 1105)[6] Wang J.L.,Shi Z.X.,Ge Y.,Wang Y.,Fan J.C.,Yin J.,Mater.Chem.Phys.,2012,136(1),43 50[7] Pramoda K.P.,Hussain H.,Koh H.M.,J.Polym.Sci.,Polym.Chem.Part A ,2010,48,4262 4267[8] Pham V.H.,Dang T.T.,Hur S.H.,Kim E.J.,Chung J.S.,ACS Appl.Mater.Interf .,2012,4(5),2630 2636[9] Wang J.,Hu H.,Wang X.,Xu C.,Zhang M.,Shang X.,J.Appl.Polym.Sci .,2011,122,1866 1871[10] Aldosari M.A.,Othman A.A.,Alsharaeh E.H.,Molecules ,2013,18,3152 3167[11] Vallés C.,Kinloch I.A.,Young R.J.,Wilson N.R.,Rourke J.P.,Composites Science and Technology ,2013,88,158 164[12] Hummers W.S.,Offeman R.E.,J.Am.Chem.Soc .,1958,80(6),1339[13] Zhou X.Z.,Huang X.,Qi X.Y.,Wu S.X.,Xue C.,Boey F.Y.C.,Yan Q.Y.,Chen P.,Zhang H.,J.Phy.Chem.Lett.,2009,113,10842 108460742高等学校化学学报 Vol.35[14] Li Y.L.,Kuan C.F.,Chen C.H.,Mater.Chem.Phys.,2012,134(2/3),677 685[15] Yang J.T.,Yan X.H.,Wu M.J.,J.Nanopart Res .,2012,14,717 726Preparation of FGO /PBMA Composites with Improved Thermal Stability †ZHANG Danfeng 1,2*,FAN Louzhen 3*,GUO Ruihua 3,FAN Zetan 3(1.College of Chemistry ,Liaoning University ,Shenyang 110036,China ;2.College of Science ,Heilongjiang Bayi Agricultural University ,Daqing 163319,China ;3.Department of Chemistry ,Beijing Normal University ,Beijing 100875,China )Abstract Graphene oxide (GO)nanosheets was modified by 3-triethoxysilylpropylamine (KH550)to get functional graphene oxide(FGO).Then FGO was reacted with butyl methacrylate(BMA)via in situ polymeri-zation with the initiator AIBN in N 2.The prepared functional graphene oxide /polybutylmethacrylate (FGO /PBMA)composites were characterized by Fourier transform infrared spectroscopy(FTIR),X-ray photoelectron spectrometry(XPS),scanning electron microscope(SEM)and thermal gravimetric analysis(TGA).The ther-mal stability of the FGO /PBMA composites were significantly enhanced compared with that of neat PBMA.The maximum mass loss temperature increased from 267ħfor neat PBMA to 339ħfor FGO /PBMA composites.Meanwhile the reaction mechanism was also discussed.Keywords Graphene oxide /polybutylmethacrylate(FGO /PBMA);in situ Polymerization;Thermal stability;Reaction mechanism (Ed.:S ,Z ,M )†Supported by the National Natural Science Foundation of China(Nos.21073018,21233003).1742 No.11 张丹凤等:氧化石墨烯/聚甲基丙烯酸丁酯复合材料的热稳定性氧化石墨烯/聚甲基丙烯酸丁酯复合材料的热稳定性作者：张丹凤， 范楼珍， 郭瑞华， 樊择坛， ZHANG Danfeng， FAN Louzhen， GUO Ruihua ， FAN Zetan作者单位：张丹凤,ZHANG Danfeng(辽宁大学化学院，沈阳110036; 黑龙江八一农垦大学理学院，大庆163319)， 范楼珍,郭瑞华,樊择坛,FAN Louzhen,GUO Ruihua,FAN Zetan(北京师范大学化学学院,北京,100875)刊名：高等学校化学学报英文刊名：Chemical Journal of Chinese Universities年，卷(期)：2014(11)引用本文格式：张丹凤.范楼珍.郭瑞华.樊择坛.ZHANG Danfeng.FAN Louzhen.GUO Ruihua.FAN Zetan氧化石墨烯/聚甲基丙烯酸丁酯复合材料的热稳定性[期刊论文]-高等学校化学学报 2014(11)。
- 1、下载文档前请自行甄别文档内容的完整性，平台不提供额外的编辑、内容补充、找答案等附加服务。
- 2、"仅部分预览"的文档,不可在线预览部分如存在完整性等问题,可反馈申请退款(可完整预览的文档不适用该条件!)。
- 3、如文档侵犯您的权益，请联系客服反馈,我们会尽快为您处理(人工客服工作时间：9:00-18:30)。
ORGANIZATION
President
General Manager Vice President Project Manager
PCB Ink Div. Sales Dept.
Industrial Paint Div.
R&D Div.
Administration Div.
QA Dept. Product Inspections
g
ou
ou
ng
ng Gu an
n
uH a
ng Ha
n
Ku nS ha
un gS ha
uJ ian
Su Zh
gZ h
ha
nZ
Zh
Sh a
Gu an
Do
Sh e
Ch
W
Ot
he
r
i
i
Increasing rate of clients
20.0% 18.0% 16.0% 14.0% 12.0% 10.0%
KING HOUSE PAINT FACTORIES
Jiangsu Factory
Taipei Factory Dong Guan Factory
DONG GUAN FACTORY •Established in 1991 •Investment: US$ 1.5 millions •Lot Size: 13,500 m2 •Number of Staff: 70 •Capacity: about 20 tons per day
Spetrophotometer Basket Bead Mill Gloss meter Thickness meter Hardness meter Bending tester Impact tester Oven Abrasion tester Viscometer Minolta Sphere Multi-angle X-Rite For Lab.
PRODUCT RANGE
I. Solvent borne Industrial Paints • Plastic coatings (one/two –component)
• • Substrates:plastic and metal surface Product Series :ABS, PU, etc.
Human Resource Dept. Accounting Dept. Import/Ex port Dept. Warehouse Administration
Packing Color Matching
Packing
Employees’ Background
Bachelor degree
18.8%
Quality Control
Viscosity Gloss Adhesion Hardness Impact Bending Solvent Resistance Color Identification
Color Management
Factor affecting color matching Condition of color matching DIGITAL COLOR Spectrophotometer
JIANGSU FACTORY
JIANGSU FACTORY
Jiangsu Equipments
Bead Mill Dissolver Tank (small) (Medium) (5 hp) (10 hp) (2 ton) (1 ton) (0.6 ton) (0.4 ton) (0.1 ton) (10 gal) 1 5 6 4 3 6 4 3 6 3
Customer Name UNEEC CHENMING IND. CORP. KARRIE INDUSTRIAL CO. LTD. G-TECH PLANT LITE-ON ENCLOSURE INC. TOP VICTORY ELECTRONICS CHENBRO COMPUTER (DONG GUAN) JING MOLD PLASTIC ELECTRONICS CO. LTD. PROVIEW INTERNATIONAL HOLDINGS LIMITED KARRIE INDUSTRIAL CO. LTD. AIC PLASTIC(SU ZHOU)CO., LTD SU ZHOU GLOBAL LIGHTING TECHNOLOGIES TNC. SU ZHOU TONGHSIEH PLASTIC &ELECTRONIC CO.,LTD SHANG HAI TAI YUAN PLASTIC CO.,LTD KUNSHAN LIUERFENG PLASTIC ELECTRON CO.,LTD SHANGHAI YINGJI ELECTRONIC PLASTIC CO.,LTD HX PLASTIC INDUSTRY CO.,LTD I-SHENG ELECTRIC WIRE & PLASTIC(KUNSHAN)CO.,LTD MINGSHENG (WUJIANG) PLASTIC CO.,LTD TA SUN CO.,LTD HWA-DENG ELECTRIC CHEMICAL(SUZHOU) CO.,LTD GREEN POINT (SUZHOU) TECHNOLOGY CO.,LTD End User IBM,Acer,Dell,Quanta Legend,NEC HP Fujis Legend,Founder,Haier,NEC IBM Legend,Founder Legend,TCL Legend,NEC Acer,Philips,Tatung Acer Philips,K-TRONICS,Dell Legend SONY,TCL Logitech Logitech,Kodak MICROTEK NEC,TATUNG ARIMA,ASUS,Leon Tien-Han Motorola,Alcatel
Substrates:metal surface Product Series:PE, AC, ME, etc.
Location of major clients
45% 40% 35% 30% 25% 20% 15% 10% 5% 0% 3% 7% 4% 2% 2% 9% 2% 15% 17% 39%
HISTORY
1998 BSI ISO 9002 certified. Developed epoxy insulating material mainly for electrical components. 1999 Added PCB ink department. 2000 Expanded production capacity and be able to produce more than 20 tons of paints per day. 2001 Jiangsu factory established. Mainly produces plastic paints for cellular phones, digital cameras, notebooks, mouse, and other computer peripherals. (It takes about 45 minutes to Shanghai.)
Single-angle (Minolta CM-2600d) Multi-angle (X-Rite MA 68II)
Color Paste Control Production Control
Solid Color (Spectramagic) Effect Color (QA-2000)
Sales Dept. Manufacturing Dept.
Color Development Dept.
Purchasing Dept.
Manufacturing Dept. Production
Pre-mixing & dispersing
Quality & Production enhancements
DONG GUAN FACTORY
DONG GUAN FACTORY
Dong Guan Industrial Paint Equipments Bead Mill Triple Roll Mill Basket Bead Mill Dissolver Tank (small) (Medium) (large) (12 inch) (5 hp) (10 hp) (15 hp) (3 ton) (1 ton) (0.7 ton) 1 4 1 1 2 1 3 4 4 4 9
17.2% 15.6%
Hale Waihona Puke 8.1%8.0% 6.0% 4.0% 2.0% 0.0%
1999
2000
2001
Order Statistics
9% 3% 22%
14%
< $300 < $1,000 < $2,000 < $4,000 < $6,000 in US dollar
52%
Major Clients
Equipments
JINAGSU FACTORY •Established in 2001 •Investment: US$ 1.5 millions •Lot Size: 4,400 m2 •Number of Staff: 35 •Capacity: more than 10 tons per day
•
• •
Soft coatings (leather/suede –like)
Substrates:plastic and metal surface Product Series :KRC, KSC, etc.
•
• •
Baking Systems (polyester/acrylic/alkyd)