Hypomethylation of glycine dehydrogenase promoter in peripheral blood mononuclear cells is a new diagnostic marker of hepatitis B virus-associated hepatocellular carcinoma

Background: Glycine dehydrogenase(GLDC) plays an important role in the initiation and proliferation of several human cancers. In this study, we aimed to detect the methylation status of GLDC promoter and its diagnostic value for hepatitis B virus-associated hepatocellular carcinoma(HBV-HCC). Methods: We enrolled 197 patients, 111 with HBV-HCC, 51 with chronic hepatitis B(CHB), and 35 healthy controls(HCs). The methylation status of GLDC promoter in peripheral mononuclear cells(PBMCs) was identified by methylation specific polymerase chain reaction(MSP). The mRNA expression was examined using real-time quantitative polymerase chain reaction(q PCR). Results: The methylation frequency of the GLDC promoter was significantly lower in HBV-HCC patients(27.0%) compared to that in CHB patients(68.6%) and HCs(74.3%)( P < 0.001). The methylated group had lower alanine aminotransferase level( P = 0.035) and lower rates of tumor node metastasis(TNM) Ⅲ/Ⅳ( P = 0.043) and T3/T4( P = 0.026). TNM stage was identified to be an independent factor for GLDC promoter methylation. GLDC mRNA levels in CHB patients and HCs were significantly lower than those in HBV-HCC patients( P = 0.022 and P < 0.001, respectively). GLDC mRNA levels were significantly higher in HBV-HCC patients with unmethylated GLDC promoters than those with methylated GLDC promoters( P = 0.003). The diagnostic accuracy of alpha-fetoprotein(AFP) combined with GLDC promoter methylation for HBV-HCC was improved compared with that of AFP alone(AUC: 0.782 vs. 0.630, P < 0.001). In addition, GLDC promoter methylation was an independent predictor for overall survival of HBV-HCC patients( P = 0.038). Conclusions: The methylation frequency of GLDC promoter was lower in PBMCs from HBV-HCC patients than that from patients with CHB and HCs. The combination of AFP and GLDC promoter hypomethylation significantly improved the diagnostic accuracy of HBV-HCC.

Performance in Survival,Development and Reproduction of Aphis glycines Matsumura Virginoparae at High Temperatures

The soybean aphid,Aphis glycines Matsumura,is an important pest of soybean,which is native to Asia.In this study,A.glycines fed on soybean(AgFS)and A.glycines fed on wild soybean(AgFW)were reared at 25℃,27℃,29℃,31℃,33℃and 35℃,respectively,and some of the life parameters were determined.At temperature ranging from 25℃to 31℃,nymphs of AgFS and AgFW all developed into adults successfully.Only a few nymphs of AgFS and AgFW developed into adults at 33℃and no nymphs could develop into adults at 35℃.Lifespan,fecundity and body size of AgFS and AgFW adults all decreased gradually with temperatures increasing from 25℃to 33℃.At 25℃,the intrinsic rate of increase of AgFS was as big as that at 27℃,which was smaller than that at 29℃,but was bigger than that at 31℃.Intrinsic rate of increase of AgFW decreased gradually with temperatures increasing from 25℃to 31℃.Nymph stage duration of AgFW was longer than or as long as that of AgFS;adult lifespan of AgFW was shorter than or as long as that of AgFS.Adult fecundity,intrinsic rate of increase and adult body size of AgFW were all smaller than or as big as those of AgFS.It showed that AgFS and AgFW both survived and developed well at temperature ranging from 25℃to 31℃,and AgFW was more adaptive to low temperatures.These results were important to study the adaptability of A.glycines to high temperatures and for predicting its dynamics in the temperature keeping rising region.

A novel procedure for identifying a hybrid QTL-allele system for hybrid-vigor improvement, with a case study in soybean(Glycine max)yield

“Breeding by design” for pure lines may be achieved by construction of an additive QTL-allele matrix in a germplasm panel or breeding population, but this option is not available for hybrids, where both additive and dominance QTL-allele matrices must be constructed. In this study, a hybrid-QTL identification approach, designated PLSRGA, using partial least squares regression(PLSR) for model fitting integrated with a genetic algorithm(GA) for variable selection based on a multi-locus, multi-allele model is described for additive and dominance QTL-allele detection in a diallel hybrid population(DHP). The PLSRGA was shown by simulation experiments to be superior to single-marker analysis and was then used for QTL-allele identification in a soybean DPH yield experiment with eight parents. Twenty-eight main-effect QTL with 138 alleles and nine QTL × environment QTL with 46 alleles were identified, with respective contributions of 61.8% and 23.5% of phenotypic variation. Main-effect additive and dominance QTL-allele matrices were established as a compact form of the DHP genetic structure. The mechanism of heterosis superior-to-parents(or superior-to-parents heterosis, SPH) was explored and might be explained by a complementary locus-set composed of OD+(showing positive over-dominance, most often), PD+(showing positive partial-to-complete dominance, less often) and HA+(showing positive homozygous additivity, occasionally) loci, depending on the parental materials. Any locus-type, whether OD+, PD + and HA+, could be the best genotype of a locus. All hybrids showed various numbers of better or best genotypes at many but not necessarily all loci, indicating further SPH improvement. Based on the additive/dominance QTL-allele matrices, the best hybrid genotype was predicted, and a hybrid improvement approach is suggested. PLSRGA is powerful for hybrid QTL-allele detection and cross-SPH improvement.

大豆对胞囊线虫(Heterodera glycines Ichinohe)1号和4号生理小种抗性的遗传分析

大豆胞囊线虫(Heterodera glycinesIchinohe)是我国大豆的全国性主要病害之一。1号和4号生理小种是黄淮地区的优势小种。以Essex×ZDD2315、Peking×ZDD2315、PI88788×ZDD2226、Peking×ZDD2226的P1、P2、F1、BC1F2为材料,用主基因+多基因混合遗传模型分析大豆对胞囊线虫1号和4号生理小种抗性的遗传机制。结果表明,ZDD2315、ZDD2226对1号生理小种的抗性受主效基因控制,未发现多基因效应,且与Peking存在相同的抗病基因;抗性遗传表现组合特异性,Essex×ZDD2315组合为3对加性主基因遗传模型,主基因遗传率72.02%,PI88788×ZDD2226组合为2对显性上位主基因遗传模型,主基因遗传率62.33%。对4号生理小种的抗性为主基因+多基因混合遗传模型,Essex×ZDD2315、Peking×ZDD2315、PI88788×ZDD2226等3个组合为3对主基因+多基因遗传模型,主基因遗传率分别为67.76%7、2.46%和53.25%,多基因遗传率分别为24.48%、21.31%和35.77%;Peking×ZDD2226表现为2对主基因遗传模型,主基因遗传率45.40%。抗性基因表现为隐性,育种上可以在早代选择。培育多抗品种应以抗4号生理小种为主要目标进行基因聚合。

Structural characterization of calcium glycinate,magnesium glycinate and zinc glycinate

Metal glycinate chelates are formed by glycine and metal compounds through chemical reactions.Calcium glycinate,magnesium glycinate and zinc glycinate are kinds of new-type and ideal nutrient supplements,which have satisfactory physico-chemical properties and bioactivities.They are important for prophylaxis and treat metal deficiency.The structural characterization shows that the metal ion is bonded to the amino and carboxyl group to form two five-membered rings.This paper mainly studies the structure characterization of the metal chelated glycinates by their solubility,infrared spectrum,thermal analysis,mass spectrometry,polycrystal difrac-tion,the metal contents and glycine contents of calcium glycinate,magnesium glycinate and zinc glycinate.

干旱和复水对大豆(Glycine max)叶片光合及叶绿素荧光的影响

选用河南省大面积种植的大豆品种豫豆29作为实验材料,通过研究逐步干旱和旱后复水条件下大豆叶片光合、叶绿素荧光等指标随土壤水分的动态变化规律,以期为大豆的水分高效利用提供理论依据。研究发现,在土壤相对含水量高于46.5%时,虽然随着土壤相对含水量的下降,豫豆29仍可以保持它的叶片水分状态;豫豆29的叶片净光合速率在土壤水分中等条件下最大,在土壤相对含水量为64.3%时,它比对照组高出11.2%(P<0.01);在实验的第3d,处理组的土壤相对含水量降为46.5%,叶片水势与对照组相比降低了7.2%(P>0.05),净光合速率为对照组的89.6%(P<0.05),但气孔导度却迅速下降为对照组的44.7%(P<0.01),这说明与叶片的光合和水分状况相比,豫豆29的气孔对土壤水分的匮缺更加敏感。复水后,豫豆29叶片的水势、净光合速率、气孔导度和叶绿素荧光等值都可以得到迅速的恢复,并在实验的最后接近对照组的水平,这表明豫豆29的叶片光合在水分胁迫解除后有迅速恢复的能力。

大豆属Glycine亚属植物花粉形态研究

对大豆属Glycine亚属中10个种的花粉进行了光学显微镜和扫描电镜的观察.发现多倍体材料的花粉大于二倍体的.并发现Glycine亚属诸种花粉的形状、萌发孔、外壁纹饰均存明显差异.

外源茉莉酸对大豆中异戊烯黄酮Glyceollins及其前体Glycinol累积的影响

以中黄13号大豆种子为材料,取生长8天的健康子叶,在其表面挖出约2×2mm的伤口,将50μl不同浓度的茉莉酸溶液和1mM的硝酸银溶液滴在伤口处,黑暗中(25℃)保温不同时间后取样,经甲醇提取,用高压液相色谱(HPLC)测定其中异戊烯化异黄酮glyceollins及其前体glycinol 的累积情况.结果显示,健康子叶中没有glyceollins和glycinol的累积,1mM硝酸银比茉莉酸具有更强的诱导glyceollins和glycinol累积的能力,glyceollins的累积与茉莉酸浓度呈负相关.在各种处理中,glyceollinⅠ、Ⅱ、Ⅲ所占比例分别为:50%～80%、10%～40%和10%～20%,但是glyceollin三种异构体的比例随保温时间的延长而有所变化,glyceollin Ⅰ比例逐渐下降,glyceollin Ⅱ所占比例逐渐升高,在1mM硝酸银处理的子叶中表现更为明显.

野生大豆(GlycinE Soja)抗蚜鉴定技术方法研究初报

目测分级鉴定野生大豆对大豆蚜的抗性是一种简便、有效的方法。被鉴定材料扣网接蚜可以大大提高蚜虫的发生量及鉴定结果的准确性。接虫虫源可以在栽培大豆上预先繁殖。野生大豆苗期及开花期对蚜虫危害的反应基本一致。

Glycine attenuates myocardial ischemia-reperfusion injury by inhibiting myocardial apoptosis in rats

Glycine is a well-documented cytoprotective agent.However,whether it has a protective effect against myocar-dial ischemia-reperfusion injury in vivo is still unknown.By using an open-chest anesthetized rat model,we found that glycine reduced the infarct size by 21% in ischemia-reperfusion injury rats compared with that in the vehicle-treated MI/R rats.The left ventricular ejection fraction and fractional shortening were increased by 19.11% and 30.98%,respectively,in glycine-treated rats.The plasma creatine kinase levels in ischemia-reperfusion injury rats decreased following glycine treatment.Importantly,administration of glycine significantly inhibited apoptosis in post-ischemia-reperfusion myocardium,which was accompanied by suppression of phosphorylated p38 mitogen-activated protein kinase and c-Jun NH2-terminal kinase,as well as the Fas ligand.These results suggest that gly-cine attenuates myocardial ischemia-reperfusion injury in vivo by inhibiting cardiomyocytes apoptosis.

Sampling Survey and Identification of Races of Soybean Cyst Nematode (Heterodera glycines Ichinohe) in Huang-Huai Valleys

Soybean cyst nematode （SCN Heterodera glycines Ichinohe） is one of the most important nationwide soybean diseases in China. A total of 38 soil specimens or locations in the area was sampled and tested for SCN races during 2001-2003 for the inspection of race distribution in Huang-Huai Valleys. A map of race distribution was constructed according to the data from both the present study and the published reports cited. Three areas, namely, the area of southeast to Jinan in Shangdong Province; the area of northern Henan Province and its border region to south of Hebei Province; and the area of Luohe, Zhoukou of Henan Province and Fuyang of Anhui Province mainly infested with Race 1 were identified. Race 4 was predominant in Shanxi Province, Beijing and the adjacent area of Henan, Shandong, and Anhui provinces, and the delta of Huanghe River in Shandong Province. Race 2 was mainly found in Liaocheng, Dezhou of Shangdong Province and Shijiazhuang of Hebei Province, and Jiaozuo and Huojia of Henan Province. Race 7 was distributed in the west part of Jiaodong Peninsula of Shandong Province and Kaifeng, Huaxian, Wenxian of Henan Province. Race 5 was found and scattered in Hebei and Henan Province. Race 9 was found in Shangqiu of Henan Province, which was reported for the first time in China. It can be seen that Race 1 and Race 4 were the two predominant races in Huang-Huai Valleys, and that research should focus on developing resistant cultivars of these races. There might exist other races in an area with some predominant races. The race substitution in the past decade was not obviously found, therefore, the results should be meaningful to future breeding for resistance to SCN in Huang-Huai Valleys.

Green Synthesis, Composition Analysis and Surface Active Properties of Sodium Cocoyl Glycinate

Cocoyl glycine was synthesized directly from coconut oil and sodium glycinate in yield of 87%. IR, ESI-MS and LC/MS analysis results showed that cocoyl glycine consists of a series of acyl glycines with different acyl chain lengths, and the proportion of each component was consistent with the composition of fatty acids derived from coconut oil. The surface tension and the Critical Micelle Concentration (CMC) of corresponding sodium salt were evaluated. The result showed that sodium cocoyl glycinate reduced the surface tension of water to a minimum value of approximately 33 mN·m-1 at CMC of 0.21 mmol·L-1.

大豆中Glycinol的分离与鉴定

Glycinol是大豆中主要的植物抗毒素和植物雌激素glyceollins的直接前体,是研究大豆中异戊烯转移酶必需的重要底物。该研究以新鲜大豆为原料,采用硝酸银诱导大豆子叶使glycinol在其中累积,再用甲醇提取,经制备高效液相色谱分离获得单一成分;经液相色谱/四极杆-飞行时间串联质谱及紫外光谱鉴定,并与标准品对照,确定所获得的单一成分为glycinol。采用该分离方法可从390g鲜重的大豆中分离得到100mg高纯度的glycinol。该研究提供了一种分离纯化和较大量制备glycinol的方法。

Identification of Co-dominant SSR Markers Associated with Genes Controlling α'- and cr-subunit-null β-conglycinin Phenotypes in Soybean (Glycine max (L.) Merr.)

Studies have shown that the three subunits of β-conglycinin are the main potential allergens of soybean sensitive patients.And β-conglycinin has adverse effects on nutrition and food processing.So solation and production of lines with lowerβ-conglycinin content has been the focus of recent soybean breeding projects.Soybean lines with deficiency in one or all subunits of β-congIycinin have been obtained.An effective and rapid system to identify such mutations will facilitate genetic manipulation of the β-conglycinin subunit composition.Here,two segregating F2 populations were developed from crosses between Cgy-1/cgy-1(CC),anα'-lacking line(△α'),and DongNong 47(DN47),a wild-type(Wt)Chinese soybean cultivar with normal globulin components,and Cgy-2/cgy-2(CB),an a-lacking line(△α),and DN47.These populations were used to estimate linkage among the egy-1(conferring α'-null)and cgy-2(α-null)loci and simple sequence repeat(SSR)markers.Seven SSR markers(Sat_038,Satt243,Sat_307,Sat_109,Sat_231,Sat_108 and Sat_190)were determined to co-scgregate with cgy-1,and six SSR markers(Satt650,Satt671,Sat_418,Sat_170,Satt292 and Sat_324)co-segregated with cgy-2.Linkage maps being composed of seven SSR markers and egy-1 locus,and six SSR markers and the cgy-2 locus were then constructed.It assigned that the egy-1 gene to chromosome 10 at a position between Sat_307 and Sat_231,and the cgy-2 gene to chromosome 20 at a position between Satt650 and Satt671.These markers should enable map-based cloning of the egy-1 and cgy-2 genes.For different subunit-deficiency types[α'-null,α-null and(α'+α)-null types],the two sets of SSR markers could also detect of polymorphism between three normal cultivars and seven related mutant lines.The identification of these markers is great significance to the molecular marker-assisted breeding of soybean/9-conglycinin subunits.

铜、砷及其复合污染对黄豆(Glycine max)影响的初步研究

通过水培实验研究了Cu、As单一及复合污染对黄豆种子萌发、幼苗生长及部分酶活性的影响 .结果表明 ,Cu、As污染明显抑制黄豆种子萌发、幼苗生长 ,对黄豆种子萌发时的呼吸强度、蛋白酶活性有显著的抑制作用 ,且随着Cu、As浓度的增加 ,抑制作用增强 ,呈负相关 ;而POD活性则随污染物浓度的增加而增加 ,呈正相关 .Cu、As污染共同存在时 ,随二者投加比例不同出现不同程度的拮抗效应 。

利用中国秋大豆(Glycine max(L.) Merr)筛选SSR核心位点的研究

选择中国秋大豆为试验材料 ,对基因组DNA进行SSR标记筛选和鉴定。经过 2 0 0个位点在琼脂糖胶上初筛和 96个位点在变性聚丙烯酰胺胶上复鉴 ,选出 6 0个位点 ,这些位点具有以下特点 :(1)分布在大豆 2 0个整合遗传连锁群 ,相邻位点间平均遗传距离在 5 0cM左右。除连锁群C2 、O上分别有 5个位点 ,G、K、M上分别有 2个位点外 ,其余 15个连锁群均分布有 3个位点 ;(2 )与 96个位点在 80份秋大豆种质检测到种质间遗传关系达到极显著相关 (r =0 .910 ) ;(3)在 80份秋大豆初选核心种质中表现出较高多态性 ,平均每个位点等位变异数为 9.3,多态性信息含量 (PIC)值为 0 .773;(4)在检测的秋大豆绝大多数种质基因组中 ,均为单一拷贝的位点 ,具有较高特异性 ;(5 )在相同的PCR扩增条件下 ,同一位点不同等位变异间易于识别且扩增强度较为一致。这套SSR核心位点的确定为中国大豆核心种质的构建奠定了基础。

不同茬口条件下的作物根渗出物对大豆胞囊线虫(Heterodera glycines)卵孵化影响

取北方5种作物轮作茬口和休闲土壤种植火豆、玉米、小麦、亚麻和甜菜,出苗后22 d制备作物的根渗出物,观察其对大豆胞囊线虫卯孵化的影响。试验表明,麦豆麦迎茬的甜菜根渗出物在后期大豆胞囊线虫(SCN)的卵孵化率最高,豆麦米轮作茬的大豆根渗出物9 d后SCN的卵孵化率明显高于其它作物,米豆米茬口的玉米根渗出物SCN的卵孵化率较高,豆麦豆迎茬甜菜根渗出物SCN的卵孵化率明显高于所有供试作物,12年大豆连作茬甜菜从一开始卵孵化率就高于对照和其它4种作物,休闲区的5种作物根渗出物也有对SCN卵孵化的刺激作用。

中国栽培大豆(Glycine max (L.) Merr.)微核心种质的群体结构与遗传多样性

【目的】评价中国栽培大豆微核心种质的群体结构和遗传多样性水平,为拓宽大豆遗传基础、发掘优异基因、改良大豆品种提供理论依据。【方法】利用大豆20个连锁群上的100个SSR位点,对来自全国28个省补充完善的248份栽培大豆微核心种质进行SSR遗传多样性及群体结构分析;采用PowerMarker Version 3.25软件统计等位变异数、平均等位变异数、多态性信息量(PIC值)及亚群特有等位变异数等参数;基于遗传距离建立了栽培大豆微核心种质的无根Neighbor-Joining树;用Structure2.2软件对微核心种质的群体结构进行评价。【结果】100个SSR位点在248份材料中共检测出等位变异1460个,每个位点变异范围为2—33个,平均为14.6个,每个位点PIC值变异范围为0.158—0.932,平均为0.743。基于模型的群体结构分析显示,依据LnP(D)无法判断最佳K值(群组数),但通过计算系数ΔK发现,K=3为微核心种质的最佳群体结构。结合种质的生态类型及品种类型分析发现,地理来源相同的种质具有聚在一起的倾向,但来源相同的种质也有分在不同组的情况。不同生态类型及品种类型间均存在较多的互补等位变异和特有等位变异。【结论】中国栽培大豆微核心种质具有丰富的遗传多样性,可以用来拓宽大豆品种遗传基础;不同生态类型及品种类型间存在较多的互补及特有等位变异,是种质创新及品种改良的物质基础;栽培大豆微核心种质存在明显的群体结构,为微核心种质在育种中的直接或间接利用提供了理论依据。

高盐碱胁迫下野生大豆(Glycine soja)体内离子积累的差异

在总含量3%高盐碱原土盆栽条件下,对搜集于津唐盐碱地895份野生大豆材料进行全生育期耐盐碱性鉴定。通过测定203株凋亡植株及收获植株茎、叶中Na+、Cl-、K+、Ca2+、Mg2+的含量,分析高盐碱胁迫下野生大豆植株离子含量的分布及野生大豆植株凋亡和成熟时体内离子积累程度,并探讨耐盐碱型野生大豆的耐性机制。结果表明,野生大豆植株在营养生长期间,盐碱胁迫致死植株茎、叶无机离子含量在不同存活时间组间并未达显著水平,Na+和Cl-积累达到一定含量即出现凋亡现象,致死植株茎中Na+和Cl-离子范围分别为3.239%～4.682%和4.639%～6.328%,叶中分别为1.754%～2.349%和4.126%～5.073%;能够存活到成熟的耐盐碱型野生大豆植株茎叶Na+和Cl-含量存在低、中、高3种类型。高耐型野生大豆茎、叶平均Na+和Cl-含量显著低于低耐盐型,且茎中K+和叶中Ca2+和Mg2+含量较高。在高耐型野生大豆植株茎叶中也存在Na+和Cl-离子含量高水平和低水平两种类型,推测野生大豆可能存在两种耐盐碱机制,其一为高耐受性,其二为低吸收性。

大豆遗传图谱的构建和抗胞囊线虫(Heterodera glycines Ichinohe)的QTL分析

大豆胞囊线虫1号和4号生理小种是黄淮地区的优势小种，ZDD2315是我国特优抗源。本文旨在定位ZDD2315对1号和4号生理小种抗性的QTL。试验以Essex为母本，ZDD2315为父本和轮回亲本，创建了一个包含114个单株的Bc。群体。采用250个SSR标记和1个形态标记通过MAPMAKER3．0构建了包含25个连锁群的遗传图谱，覆盖大豆基因组2963．5cM，平均每个连锁群上10．0个标记，标记平均间距11．8cM。采用Win QTL Cartographer Version 2．5复合区间作图法（CIM）检测到3个抗1号小种的QTL；其中rhgR1-1和rhgR1—2位于G连锁群的Sat_210～Sat_168和Sat_168～Sat_141区间，贡献率分别为22．4％和21.8％；rhgR1-3位于D2连锁群的Satt672～Satt413区间，贡献率6．2％；rhgR1-1和rhgR1—3分别与Sat_210和Satt672共分离。5个QTL与抗4号生理小种有关；其中rhgR4—1和rhgR4—-位于G连锁群的Satt275～Sat_210和Sat_168～Sat_141区间，贡献率分别为22．8％和28．9％；rhgR4—3和rhgR4—4位于H连锁群Satt442～Sat401和Sat_334～Satt181区间，贡献率分别为12．0％和10．5％；rhgR4—5位于L连锁群Satt652～Sat_301区间，贡献率5．9％；吨职4—2和rhgR4—5分别与Sat_168和Satt652共分离。不同遗传体系控制ZDD2315对1号和4号小种的抗性。抗1号和4号生理小种的主要QTL位于G连锁群的相近区段，且具有较大贡献率，通过标记辅助选择有可能育成兼抗两小种的品种。