部分梨品种基因组DNA不同提取方法的比较

以不同梨品种为材料,分别采用改良CTAB法、改良SDS法、SDS-CTAB和高盐低pH值法对梨基因组DNA进行了提取.结果表明：采用改良CTAB法在提取各种梨基因组DNA过程中表现最好且纯度最高;SDS法提取的基因组DNA产率最高,但纯度较低;高盐低pH值法产率和纯度均最低.     

节瓜基因组DNA提取方法比较研究

以节瓜的成熟干种子、子叶和不同发育阶段的真叶为材料,选用尿素提取法、高盐低pH值法、SDS法和CTAB法等4种DNA提取方法进行比较。结果表明:不同节瓜材料均可以提取到基因组DNA,其中子叶和第1片真叶的提取效果最好;尿素提取法提取种子DNA最为简便快捷,CTAB法提取叶片能得到纯度较高的基因组DNA。经PCR检验,干种子和子叶提取的基因组DNA都可用于PCR扩增。     

几种三华李基因组DNA提取方法的比较研究

以CTAB法、改良CTAB法、SDS法、改良SDS法、改良陈大明法、高盐低pH值法、SDSCTAB法和氯化苄法等8种目前常用植物基因组DNA提取方法为基础,通过采用异丙醇和无水乙醇进行2次常温短暂沉淀、提高离心力等多种去杂质措施进行适当改进,对它们用于三华李基因组DNA提取的效果进行比较。结果表明,8种方法中以CTAB法最优,改良CTAB法其次,高盐低pH值法稍逊,其他5种方法则不适合于三华李基因组DNA的提取;CTAB法不仅适合于早食李不同季节(4月、8月和12月)和不同成熟度叶(未展开嫩叶、展开嫩叶、成熟叶和老叶)基因组DNA的提取,也适合于华蜜大蜜李、白脆鸡麻李、大鸡麻李、中蜜李、小蜜李、串珠李、从化三华李、红线李、香蕉李、中熟李和软枝三华李等11个三华李品种嫩叶基因组DNA提取。     

落叶期牡丹非叶材料提取DNA的方法

以牡丹休眠时期根、茎、芽为材料,用CTAB法、SDS法、高盐低pH法进行DNA提取.结果表明:采用CTAB法、SDS法、高盐低pH法均能从不同采样期的根、茎、芽中提取出DNA,采样时期对提取效果没有显著影响;以牡丹根采用CTAB法提取,所获得的DNA产量高、质量优,是获取落叶期牡丹DNA的最佳组合.     

越橘基因组DNA的快速提取及分析

为了从富含多酚、多糖及色素的越橘叶片中提取适用于分子生物学研究的高质量基因组DNA。以越橘幼叶为实验材料,比较了CTAB、SDS、高盐低pH值3种提取方法,获得了一种以CTAB法为基础的分离高质量完整DNA的简便、快速方法。用紫外分光光度计、琼脂糖凝胶电泳、RAPD-PCR、酶切等方法对获得的DNA进行了分析,结果表明,快速CTAB法所提取的DNA产量高、质量好,完全能够满足RAPD、PCR等分子生物学实验的要求。     

辣椒基因组DNA提取方法研究

本项研究采用SDS法、CTAB法和高盐低pH法对辣椒(CapsicumannuumL.)叶片基因组DNA进行提取;紫外吸收检测法与琼脂糖凝胶电泳法对DNA的纯度进行检测。紫外吸收检测结果表明,SDS法提取的辣椒叶片DNA具有典型的天然DNA分子的标准紫外吸收光谱特点,其A260/A280在1.771～1.912之间。SDS法提取的DNA经琼脂糖凝胶电泳检测得到一条迁移率很低的整齐清晰的DNA谱带,所提取DNA的质量和产率均较高,用该法提取的辣椒DNA进行RAPD分析,DNA扩增效果较好,带形清晰、整齐,说明SDS法提取的DNA分子较为完整,能用作PCR模板来开展辣椒分子水平的研究。     

菊芋基因组DNA提取方法的比较

采用改良CTAB法、改进的高盐SDS法及两种植物基因组DNA提取试剂盒法等４种方法提取菊芋基因组DNA,并进行电泳分析、质量检测及ISSR引物扩增检测。结果表明,改良CTAB法优于其他3种方法,提取的DNA质量较好、纯度较高,能够满足ISSR-PCR扩增要求。     

甜菜干种子DNA提取的不同方法比较

分别采用SDS法、CTAB法及碱裂解法对甜菜干种子基因组DNA进行提取．并利用1％琼脂糖判断DNA的纯度,λDNA判断DNA的含量,结果表明,SDS法和CTAB法提取的甜菜基因组DNA含量较高,提取的DNA总量接近,碱裂解法提取的DNA含量较低。3种方法提取的DNA均能够满足SSR—PCR的要求,并且扩增务带没有差异,由于碱裂解法提取DNA快速和简单,因此适合大群体DNA的提取及对模板要求不高的PCR反应中,而SDS法及CTAB法适合一次性大量提取甜菜基因组DNA以及对于DNA纯度要求较高的PCR反应中。     

不同方法对枣叶片总DNA提取效果的影响

以串杆枣的成熟鲜叶为试材,分别采用-20℃、-70℃和硅胶脱水干燥3种方法保存样品,采用简易CTAB法、高盐沉淀法与高盐低pH值法分别对3种方法保存的样品及对照样(鲜叶)进行了基因组DNA提取效果的比较分析。结果表明,3种方法均能有效地从枣叶片中获得质量较高的DNA,以高盐沉淀法所得的DNA纯度最高,高盐低pH值法次之。就DNA的产量而言,高盐沉淀法的DNA产量相对较低,低于其他的2种方法,但未达到显著水平。硅胶干燥法和-70℃超低温保存的样品与鲜叶相比,所提DNA的质量和产量均无明显差异,均能满足PCR扩增要求。硅胶保存法适于野外远距离采样。     

不夜城芦荟基因组DNA提取方法的比较研究

以不夜城芦荟的幼嫩叶片为试材,分别采用SDS法和CTAB法以及简化的CTAB法对基因组DNA进行提取,并利用紫外分光光度计和琼脂糖凝胶电泳法检测其DNA的浓度、纯度及质量,以期筛选不夜城芦荟基因组DNA提取的最合适方法.结果表明:对比不夜城芦荟基因组DNA提取的3种方法,无论从提取浓度还是纯度上来看,SDS法均明显优于其它方法.     

Effects of radiation from 188Re on smooth muscle cell proliferation

AIM: Although endovascular radiotherapy inhibits neointimal hyperplasia, the exact alterations induced by β-particles irradiation remain to be elucidated. The objective of this study was to investigate the ability and the cellular mechanism of local β^-particles emission from ¹⁸⁸Re to inhibit vascular smooth muscle cells (SMCs). METHODS: The SMCs in vitro were irradiated by ¹⁸⁸Re with single doses of 2.6 Gy-25.8 Gy. The effects of β-particles on SMCs, such as effective irradiate doses, the period of inhibition for SMCs proliferation, the changes of cell proliferation rate and DNA synthesis rate, cell cycle progression and related gene expression, were investigated by cell count, [³H]-TdR incorporation, cell cycle progression analysis, cell viability and immunocytochemistry, respectivecy. RESULTS: β-particles irradiation with dose of 5.2 Gy could inhibit significantly SMCs proliferation. At dose of 20.6 Gy DNA synthesis inhibitory rate was 92%, SMCs proliferation rate was only 3%. Renoval of ¹⁸⁸Re did not abolish the inhibitory effects of β-particles on SMCs proliferation. The expression of P53 was up regulation and PCNA was down regulation after irradiation. CONCLUSION: β-particles from ¹⁸⁸ Re was significantly effective and permanent in inhibiting SMCs proliferation, and inhibitory effect was in dose-dependet manner ED50was 5 Gy, the best dose to inhibit SMCs proliferation was 20 Gy. β-particles irradiation induced SMCs to occur G₀/G₁ arrest, damaged the ability of SMCs reproliferation and led to cell clonogenic death. P53 and PCNA had regulatiory effects on SMCs proliferation after β-particles irradiation.     

Effect of L-Arg on plasma content of endothelin and expression of c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy

AIM:To study the effect of L-Arg on plasma content of endothelin (ET) and the expression of proto-oncogene c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy. METHODS: The level of c-fos mRNA were measured by in situ hybridization. The ET in plasma were measured by radioimmunoassay. RESULTS:After eight weeks of treatment with L-Arg, the expression of c-fos decreased markedly (P＜0.01). The ET content in plasma also decreased significantly by L-Arg(P＜0.01).CONCLUSION: Plasma ET content and the expression of c-fos in the left ventricle of rats with renovascular hypertensive hypertrophy could be decreased by L-Arg administration.     

Ergebnisse der Leistungsprüfungen der Süßkirschensorte ‚Stella‘ auf Gisela- und Weiroot-Unterlagen in Bulgarien

Zusammenfassung Die Leistungsprüfungen wurden im Zeitraum 1997 bis 2003 mit den Unterlagen Gisela 4 und 5, den Klonnummern 195/20 und 497/8 aus der Gisela-Serie sowie Weiroot 10, 13, 53, 72 und 158 durchgeführt. Dabei dienten Sämlinge von P1 (bulgarische Selektion aus Prunus mahaleb) als Kontrolle. Alle Unterlagen waren mit der Sorte Stella veredelt und im Dezember 1996 in der Versuchsanlage der Agraruniversität in Plovdiv, Bulgarien, im Abstand von 6 m×4,5 m gepflanzt worden. Dabei erfolgte ein Pflanzschnitt. Nach Abschluss der natürlichen Kronenentwicklung wurde jedes Jahr ein Winterschnitt vorgenommen. Der Boden wurde durch mechanische Bearbeitung offen gehalten und nach dem 4. Standjahr wurden die Baumstreifen mit Herbiziden behandelt. Die Wasserversorgung erfolgte durch eine dem natürlichen Gefälle folgende Überflutung, allerdings nicht immer zum optimalen Zeitpunkt, da keine eigene Wasserquelle zur Verfügung stand.Basierend auf den Ergebnissen bis zum Anfang des 7. Standjahres können die untersuchten Unterlagen in zwei Gruppen differenziert werden: starkwüchsig—Weiroot 10, P1 und Weiroot 13; mittelstarkwachsend bis schwachwüchsig—Gi 497/8, Gisela 4, Weiroot 53, Weiroot 158, Gi 195/20, Weiroot 72 und Gisela 5. Letztere zeichnete sich durch besondere Schwachwüchsigkeit aus. Die meisten Wurzelschosser bildeten Gisela 4, Weiroot 10 und Weiroot 13. Weiroot 53, Weiroot 72 und Weiroot 158 entwickelten deutlich weniger und P1, Gisela 5, Gi 195/20 sowie Gi 497/8 keine Wurzelschosser. Den frühesten Blühbeginn induzierte Gisela 4. Die anderen Unterlagen führten, in Abhängigkeit von den Temperaturbedingungen des jeweiligen Jahres, zu einer Verspätung der Blüte: P1 und Weiroot 10 um 1–2 Tage; Gi 497/8, Weiroot 13 und Weiroot 158 um 2–4 Tage; Weiroot 72 um 2–7 Tage; Gi 195/20 um 3–6 Tage; Weiroot 53 um 3–8 Tage und Gisela 5 um 3–10 Tage. Die Reifezeit der Früchte war bei den Bäumen auf Gisela 5 im Vergleich zu den anderen Varianten um 2–3 Tage verspätet. Gisela 5, Weiroot 72 und Gisela 4 induzierten bei der aufveredelten Sorte die höchsten Ertragsleistungen, P1 die geringsten. Bei den Bäumen auf Gisela 5 war die Fruchtgröße geringer als bei den anderen Unterlagen. Bäume auf Gisela 5 brauchen intensive Pflege. Nur wenn alle Produktionsfaktoren und kulturtechnischen Maßnahmen optimiert werden, kann das hohe Ertragspotenzial dieser Unterlage ausgeschöpft werden.     

Compositional and Functional Properties of Saskatoon Berry and Blueberry

Abstract

Saskatoon berry (Amelanchier alnifolia Nutt., Rosaceae) and blueberry (Vaccinium corymbosum L., Ericaceae) are substantially equivalent in all characteristics that are important to the consumer, including fruit color, shape, size, nutrition, texture, and uses. In addition, both fruits are native to North America and they have practically identical historical uses and known health benefits. Their composition, processing, nutritional value and metabolism, intended uses, and levels of undesirable substances are compared.     

多效唑对猕猴桃离体试管苗生长及内源激素的影响

多效唑（ＰＰ３３３）处理猕猴桃试管苗，降低了其生长强度；植株体内的ＧＡ３、ＩＡＡ和ＺＴ含量下降，ＡＢＡ的含量上升，乙烯释放率增加；并且能降低外源的ＧＡ３和ＩＡＡ促进生长的作用，而外源的ＧＡ３和ＩＡＡ又能不同程度地逆转多效唑的抑制作用，使植株恢复生长。     

Proteomic analysis between pathological scars and normal skin

AIM: To investigate and screen the sensitive proteins in the formation mechanism of pathological scars by comparing the results of differential proteomic analysis between pathological scars and normal skin.METHODS: Two-dimensional gel electrophoresis was used to detect the protein expression profiles in 8 keloid patients, 8 hypertrophic scar patients and 3 matched normal skin patients.The proteins that showed differential expression of over 4-fold change were cut and analyzed by MALDI-TOF/TOF mass spectrometry.RESULTS: A two-dimensional protein profiling comparison between pathological scars and normal skin was successfully established.On average, 2 978 spots in keloid, 2 975 spots in hypertrophic scar and 3 053 spots in normal skin were identified using gel analysis software.Compared with normal skin, there were totally 36 differentially-expressed proteins in keloid and hypertrophic scar identified from the spots of over 4-fold change, including 16 proteins in both keloid and hypertrophic scar (8 up-regulated and 8 down-regulated), 11 only in keloid (9 up-regulated and 2 down-regulated) and 9 only in hypertrophic scar (4 up-regulated and 5 down-regulated).CONCLUSION: Proteomic analysis can identify the proteins with variance of pathological scars versus normal skin, thus providing probable new clues to reveal the formation mechanism of pathological scars.     

Cryopreservation of shoot apices of hawthorn in vitro cultures originating from East Asia

The objective of this study was to establish a cryopreservation protocol for hawthorn shoot apices (Crataegus pinnatifida Bge.). Cryopreservation was carried out via encapsulation–dehydration, vitrification, and encapsulation–vitrification on shoot apices excised from in vitro cultures. We began by showing that cold-acclimation enhanced the regrowth of cryopreserved apices from 10.0 to 65.5% in encapsulation–dehydration. We then decided that the encapsulation–dehydration method was an optimal cryopreservation method for hawthorn shoot apices in terms of its high recovery after cryopreservation as well as its ease of use compared with vitrification and encapsulation–vitrification. In encapsulation–dehydration, the protocol leading to optimal regrowth was as follows: after cold-acclimation at 5 °C in the dark for 2 weeks, excised shoot tips were pretreated for 24 h at 25 °C on hormone-free Murashige and Skoog [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15, 473–497] (MS) basal medium with 0.4 mol/L sucrose, then encapsulated and precultured in liquid MS medium with 0.8 mol/L sucrose for 16 h at 25 °C. Precultured beads were dehydrated for 6 h at 25 °C in the dessicator containing 50 g silica gel to a moisture content of 15.3% (fresh-weight basis) before cryostorage for 1 h. In addition, we examined the effect of adding glycerol to both the alginate beads and loading solution to enhance regrowth after cryopreservation in encapsulation–dehydration. In the present study, it was shown that adding 0.5 mol/L glycerol resulted in high regrowth percentages (82.5–90.0%) in four Crataegus species.     

Metallothionein inhibits homocysteine-induced proliferation of rat vascular smooth muscle cells

AIM:To investigate the effect of metallothionein(MT) on proliferation of rat vascular smooth muscle cells (VSMCs) stimulated by homocysteine and its mechanism. METHODS:VSMCs proliferation was measured by [³-H]-TdR incorporation, mitogen-activated protein kinase(MAPK)activity were determined by immunoprecipitation method, the intracellular contents of MT and malondialdehyde (MDA)were assayed by -hemoglobin saturation method and TBA reaction, respectively, and lactate dehydrogenase (LDH) leakage was measured by NADH oxidation. RESULTS:Hcy(10^-6-10^-4 mmol/L) stimulated [³-H]-TdR incorporation by the VSMCs in a concentration-dependent manner. Compared with control, [³-H]-TdR incorporation in VSMCs treated with 0.1 mmol/L Hcy was increased by 4.2 fold (P＜0.01). Meanwhile, Hcy enhanced MAPK activity, MDA formation and LDH release (P＜0.01)in a concentration-dependent manner. Treatment of VSMCs with MT alone did not change above parameters, compared with control. However, MT (10^-6-10^-4 mol/L)attenuated significantly Hcy-stimulated proliferation of VSMCs (P＜0.01)in a concentration-dependent manner. And MT inhibited obviously Hcy-induced activation of MAPK activity, MDA formation and LDH release. Preincubation of VSMCs with 0.5 mmol/L ZnCl₂ for 6 h induced an increase cellular MT content by 5.7-fold (P＜0.01). The MT-overexpressed VSMCs resisted Hcy-stimulating action on MAPK activity, MDA formation and LDH leakage (P＜0.01). CONCLUSION:These results show that MT has an inhibitory effect on Hcy-induced VSMCs proliferation, and that MT could inhibit Hcy-stimulated MAPK activity and lipid peroxidation.     

Coupling past management practice and historic landscape change on John F. Kennedy Space Center,Florida

Historic landcover dynamics in a scrubby flatwoods (Tel-4) and scrub landscape (Happy Creek) on John F. Kennedy Space Center were measured using aerial images from 1943, 1951, 1958, 1969, 1979, and 1989. Landcover categories were mapped, digitized, geometrically registered, and overlaid in ARC/INFO. Both study sites have been influenced by various land use histories, including periods of range management, fire suppression, and fire management. Several analyses were performed to help understand the effects of past land management on the amount and spatial distribution of landcover within the study sites. A chi-squared analysis showed a significant difference between the frequency of landcover occurrence and management period. Markov chain models were used to project observed changes over a 100-year period; these showed current management practices being effective at Tel-4 (restoring historic landscape structure) and much less effective at Happy Creek. Documenting impacts of past management regimes on landcover has provided important insight into current landscape composition and will provide the basis for improving land management on Kennedy Space Center and elsewhere.     

XBP-01 accelerated apoptosis induced by oxidized low density lipoprotein in vascular smooth muscle cells

AIM: Previous studies performed with XBP-01 in vitro indicated that XBP-01 could inhibit vascular smooth muscle cells from being transformed into foam cell and could eliminate the atherosclerotic plaque in C57BL/6J mouse. This experiment is to investigate its mechanism of eliminating plaques in vitro. METHODS: The cultured porcine artery smooth muscle cells incubated with XBP-01 of 0.1 mg/L for 24 h after preincubated with oxidized low density lipoprotein of 15 mg/L for 72 h in vitro. The samples were analyzed by fluorescence microscope, confocal microscope system and flow cytometry. RESULTS: Apoptosis was triggered by being incubated with oxidized low density lipoprotein and this process was accelerated additionally by being incubated with XBP-01. CONCLUSION: XBP-01 can be effective in eliminating atherosclerotic plaque by accelerating the process in which oxidized low density lipoprotein induced smooth muscle cell apoptosis.